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. 2021 Jan 2;11(1):12. doi: 10.1007/s13205-020-02557-y

LncRNA HOXA11-AS regulates the proliferation and epithelial to mesenchymal transition of human skin cancer cells

Jigang Wang 1,✉, Xiuli Li 1, Hui Li 1, Xin Li 1
PMCID: PMC7778660  PMID: 33442511

Abstract

LncRNA HOXA11-AS functions as regulator of tumorigenesis of several human cancers. The present study was intended to explore its regulatory control in human skin cancer with emphasis on understanding the underlying molecular mechanism. The results showed significant (P < 0.05) upregulation of lncRNA HOXA11-AS transcript levels in human skin cancer tissues and cell lines. The knockdown of HOXA11-AS significantly (P < 0.05) inhibited the proliferation and colony formation of A375 and HMCB skin cancer cells. Flow cytometry showed that HOXA11-AS knockdown triggered arrest of the A375 and HMCB cells at G2/M check point of cell cycle by inhibiting the expression of cyclin B1. Additionally, western blot analysis showed that HOXA11-AS knockdown resulted in the deactivation of PI3K/AKT/mTOR signaling pathway. The silencing of HOXA11-AS significantly (P < 0.05) inhibited the migration and invasion of the A375 and HMCB skin cancer cells. This was also accompanied by increase in E-cadherin and decrease in N-cadherin expression. Collectively, the results indicate that lncRNA HOXA11-AS regulates the proliferation, migration and invasion of human skin cancer and may exhibit therapeutic potential in the treatment of skin cancer.

Keywords: Skin cancer, Melanoma, HOXA11-AS, MTT assay, Epithelial to mesenchymal transition, Western blotting, Cell cycle arrest

Introduction

The skin cancer or melanoma is collectively used to describe both malignant and non-malignant melanoma (Whiteman et al. 2016). The malignancy is having highest prevalence among the whitish people while the ethnic groups are least affected (Apalla et al. 2017). Also, the incidence of skin cancer is seen to increase with increasing age and average diagnosis age has been reported to be 60 years (Leiter et al. 2017). The males are seen to possess 1.5 times higher prevalence than the female population (Ribero et al. 2016). Of all skin cancers, almost 80 percent of the mortality results from the malignant melanoma (Dimitriou et al. 2018). The 5 year survival rate of malignant melanoma is less than 15% attributed to its poor prognosis and exceedingly high metastasis (Schadendorf et al. 2018). Less admirable clinical success has been achieved against the skin cancer through application of surgical procedures and chemo or radio-therapeutic approaches. Therefore, it becomes important to understand the mechanics of skin cancer at molecular level through evaluation of various regulatory aspects controlling the pathogenesis and spread of this malignancy. During recent times, it has been deduced through various research investigations that non-protein coding RNA classes including the long non-coding RNAs (lncRNAs) and micro-RNAs (miRNAs) act as crucial gene regulators to affect various biological processes like cell division, differentiation, apoptosis, invasion, migration, etc. (Hauptman and Glavac 2013; Di Gesualdo et al. 2014) Growing evidences suggest that lncRNAs function as key regulators of growth and progression of human cancers (Zhang et al. 2016; Lin and Yang 2018). Studies have shown that lncRNAs regulate the molecular mechanics of melanoma in humans (Aftab et al. 2014). For instance, the growth and metastatic behavior of melanoma cells was reported to be regulated by lncRNA GAS5 (Chen et al. 2016). The epithelial to mesenchymal transition of cancer cells has been implicated to be regulated by the function of lncRNAs (Xu et al. 2016). The epithelial to mesenchymal transition of non-small lung cancer cells was shown to be modulated by lncRNA SPRY4-IT1 whose epigenetic suppression promoted the cell proliferation and metastasis (Sun et al. 2014). The lncRNA HOXA11-AS belongs to the HOXA gene cluster of homeobox genes (Cui et al. 2017). The HOXA11-AS gene is 3885 bp in length and is located on 7p15.2 chromosomal section (Richards et al. 2015). Researchers have suggested lncRNA HOXA11-AS functions as vital prognostic biomarker and therapeutic target in many human cancers (Wei et al. 2020; Lu et al. 2018). It was shown to regulate the epithelial to mesenchymal transition of breast cancer cells (Li et al. 2017). However, its role in regulating the mechanics of epithelial to mesenchymal transition of human melanoma cells has not been studied and thus, current study was aimed at the same. The results showed that HOXA11-AS is up-regulated in skin cancer. The silencing of HOXA11-AS inhibited the proliferation of cancer cells through G2/M cell cycle arrest which was modulated through PI3K/AKT/mTOR signaling pathway. Further, HOXA11-AS was confirmed to regulate the epithelial to mesenchymal transition of cancer cells. The study suggested the prognostic and therapeutic value of HOXA11-AS against the human skin cancer.

Materials and methods

Tissue samples

After obtaining written consent, twenty-five paired cancerous and normal-adjacent skin tissues were obtained from the skin cancer patients admitted in Cangzhou Central Hospital. All those skin cancer patients who had not undergone radio- or chemo-therapy prior to the collection of the tissue samples were included in the present study (inclusion criteria). Those skin cancer patients who had undergone radio- or chemo-therapy were excluded from the study (exclusion criteria). This was done to avoid the effects of radio- or chemotherapy on the expression profile of the LncRNA HOXA11-AS. The patients were diagnosed by taking the skin biopsy samples and subsequent examination by a pathologist. The tissues were stored frozen using liquid N2. The hospital ethical guidelines were strictly followed to use the clinical specimens for the experimental purpose. The study was approved by research ethics committee of Cangzhou Central Hospital, Cangzhou, Hebei, China on 6th of June 2019 with approval number CCH/0331/2019.

Cell lines and culture conditions

The skin cancer cell lines (A431, A375, A101D, HMCB and Hf688) and the normal human skin cell line (HEMn-LP) were obtained from the Shanghai Cell Institute of Chinese Academy of Sciences. The Dulbecco's modified Eagle medium (DMEM) added with 10% FBS was used for the laboratory maintenance and culturing of all the cell lines at 37 °C with 5% CO2 in humidified incubator.

Transfection

The transfection of skin cancer cells was performed using Lipofectamine 2000 reagent (Thermo Fisher Scientific) as per the manufacturer guidelines. The knockdown anti-sense construct of HOXA11-AS (si-HOXA11-AS) and its negative control (si-NC) were designed and supplied by RiboBio biotechnology company, China.

Expression analysis

Total-RNA from the frozen tissues and cell lines was extracted with the help of TRIzol® reagent (Thermo Fisher Scientific) as per the manufacturer's protocol. Using M-MLV Reverse Transcriptase (Takara), cDNA was synthesized and then RT-PCR was performed on QuantStudio 5.0 PCR system (Thermo Fisher Scientific) with SYBR Green PCR master mix. The relative expression of HOXA11-AS was calculated with the help of 2−△△Ct method. Human β-actin served as an expressional control. The primers used were: HOXA11-AS:F, 5′-TGCCAAGTTGTACTTACT-3′; R, 5′-AGGAGTAGGAGT ATGTA-3′, and β-actin: F, 5′-CCTGGATAGC AACGTAC-3′; R, 5′-CACCTTCTACAATGAGCT-3′.

Proliferation and viability assays

The transfected skin cancer cells were seeded into 96-well plates at cellular concentration of 104 cells/well. After 0, 12, 24, 48 or 96 h growth at 37 °C, 20 μL MTT solution (1.5 g/L) was added into each well and incubation of 37 °C was prolonged again for 4 h. Afterwards, 175 μL DMSO was inoculated into each well. The OD values at 570 nm were taken using spectrophotometer and proliferation curves were drawn. The viability of transfected cancer cells was analyzed by the clonogenic assay. Briefly, the cells were added into 6-well plates at 103 cells/well. The cells were grown for two weeks at 37 °C in the humidified incubator with 5% CO2. The culture medium was replaced after every 3 and half days, the colonies were PBS washed, fixed by ethanol after 14 days, and 0.1% crystal violet solution was used for their staining. Finally, the colonies were visualized under light microscope and photographs were taken.

Flow cytometry

The skin cancer cells were transfected for 48 h with si-HOXA11-AS or its negative control si-NC. 1.5 mL of culture medium carrying 2 × 105 cells was added into 6-well plates. The transfected cells were cultured continuously for 48 h at 37 °C after which the cells were harvested and washed with PBS. The cells were again collected, suspended in pre-chilled 70% ethanol, and fixed at − 20 °C. The cells were then stained using propidium iodide (PI) solution and flow cytometry was used to analyze the mitotic phase distribution of the cancer cells.

Transwell chamber assay

The migration of the si-NC and si-HOXA11-AS transfected A375 and HMCB skin cancer cells was examined by transwell chamber method. Approximately, 105 transfected cells were cultured in the upper chamber of transwell plate which was separated from the underlying chamber by filter paper of 8 µm pore size. The lower chamber contained RPMI 1640 medium supplemented with 10% FBS. After cell culturing for 48 h at 37 °C, the filter paper was carefully removed and its upper surface was cleared from the adhering cells. The cells sticking to lower surface were washed with PBS, fixed with 4% paraformaldehyde and then stained using 0.1% crystal violet. Lastly, the cells were examined under high power light microscope to determine the cell migration. Also, the percent cell migration was calculated using random microscopic fields. The invasion of cancer cells was also determined in the same manner except for the transwell chamber plate was fitted with matrigel.

Western blotting

With the help of RIPA lysis buffer, the total cellular proteins were isolated at 4 °C. Approximately, 40 mg of proteins were separated on 10% SDS-PAGE and then blotted to nitrocellulose membrane (Millipore). The membrane was then exposed to primary antibodies at 4 °C overnight and subsequently, the exposure of secondary antibodies for 2 h was given at room temperature. ChemiDoc XRS system (Bio-Rad) was used to measure the intensity of the protein bands to analyze the concentration of respective proteins. The blotting pictures representing the protein bands were obtained.

Statistical analysis

Each experiment was carried out using three independent replicas. The data were presented as mean ± standard deviation. The statistical analysis was carried out with the help of SPSS 18.0 software. Student’s t test and one-way ANOVA were used to assess the statistical significance between two data sets. The p values < 0.05 were considered as statistically significant.

Results

HOXA11-AS is upregulated in skin cancer

The transcript levels of HOXA11-AS from different tissue specimens and cell lines were estimated using the RT-PCR method. The results suggested that HOXA11-AS was significantly up-regulated in the skin cancer tissues (Fig. 1a). Similar inference was drawn from the HOXA11-AS expression study from the cell lines. All the skin cancer cell lines (A431, A375, A101D, HMCB and Hf688) exhibited significantly higher gene expression of HOXA11-AS as compared to the normal human skin cell line, HEMn-LP (Fig. 1b). The cancer cell line (HMCB) was seen to possess the lowest HOXA11-AS expression among all the cancer cell lines. The results thus reveal that lncRNA HOXA11-AS has significant up-regulation in skin cancer.

Fig. 1.

Fig. 1

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HOXA11-AS is significantly up-regulated in skin cancer. The qRT-PCR expression study of HOXA11-AS in a clinical tissue specimens of skin cancer and normal adjacent tissues b skin cancer cell lines (A431, A375, A101D, HMCB and Hf688) and normal skin cell line (HEMn-LP). The experiments were performed in triplicate and expressed as mean ± SD (*p < 0.05)

Proliferation and viability of cancer cells markedly declines under HOXA11-AS silencing

To investigate whether HOXA11-AS regulates the growth of skin cancer cells, HOXA11-AS was silenced in A375 and HMCB cancer cells and the same was confirmed by the RT-PCR (Fig. 2a). The proliferation of cancer cells under HOXA11-AS knockdown was shown to be significantly lower as compared to the normal control cancer cells (Fig. 2b). Also, the cancer cells were seen to exhibit significantly lower colony formation under HOXA11-AS gene silencing (Fig. 3). Together, the results suggest that HOXA11-AS promotes the growth of skin cancer and its repression declines the proliferation and viability of skin cancer cells highlighting its therapeutic value.

Fig. 2.

Fig. 2

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Knockdown of HOXA11-AS restricts the proliferation of skin cancer cells. a qRT-PCR expression analysis for confirmation of transfection mediated knockdown of HOXA11-AS in A375 and HMCB cancer cells b MTT assay for estimation of proliferation of si-NC and si-HOXA11-AS transfected A375 and HMCB cells. The experiments were performed in triplicate and expressed as mean ± SD (*p < 0.05)

Fig. 3.

Fig. 3

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Knockdown of HOXA11-AS led to the loss of viability of skin cancer cells. Clonogenic assay for the determination of relative in vitro colony forming potential of si-NC and si-HOXA11-AS transfected A375 and HMCB cells. The experiments were performed in triplicate and expressed as mean ± SD (*p < 0.05)

HOXA11-AS knockdown induced mitotic cell cycle arrest in skin cancer cells

The mitosis of A375 and HMCB skin cancer cells under HOXA11-AS gene knockdown in comparison to respective control cells was studied through flow cytometric method. The relative concentration of cancer cells representing G0/G1, S and G2/M was analyzed. It was seen that the concentration of cancer cells down-regulating HOXA11-AS gene was significantly higher for G2/M phase for both A375 and HMCB cancer cells (Fig. 4a). However, no correlation could be drawn for cells representing G0/G1 and S phases of mitosis. The western blotting of cyclin B1 shown that silencing of HOXA11-AS reduced the expression of cyclin B1 protein, which is important for mitotic entry (Fig. 4b). The findings thus suggest that silencing of HOXA11-AS in skin cancer cells induced G2/M phase mitotic cell cycle arrest which was manifested as fall in proliferation and loss of cellular viabilities.

Fig. 4.

Fig. 4

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Silencing of HOXA11-AS induced G2/M mitotic cell cycle arrest in skin cancer cells. a Flow cytometric analysis of mitotic phase distribution of in si-NC and si-HOXA11-AS transfected A375 and HMCB cells b Western blots showing expression of cyclin B1 in si-NC and si-HOXA11-AS transfected A375 and HMCB cells. The experiments were performed in triplicate and actin was used as a control

PI3K/AKT/mTOR signaling pathway was inhibited under HOXA11-AS knockdown

Western blotting technique was used to examine the protein expression levels of signaling components of one of the important cell signaling pathways, PI3K/AKT/mTOR pathway. It was seen that the expression of the phosphorylated/activated PI3K, AKT and mTOR proteins drastically fall under HOXA11-AS silencing while the expression of non-phosphorylated proteins of respective signaling molecules remained unchanged (Fig. 5). Thus, the results showed that knockdown of HOXA11-AS inhibited the phosphorylation of signaling components of PI3K/AKT/mTOR signaling cascade and therefore reduced the intensity of the signaling flux in skin cancer cells.

Fig. 5.

Fig. 5

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HOXA11-AS knockdown led to blockage of PI3K/AKT/mTOR signaling pathway in skin cancer cells. Western blotting study of expression of phosphorylated (prefixed with p) and non-phosphorylated components of PI3K/AKT/mTOR signaling pathway in si-NC and si-HOXA11-AS transfected A375 and HMCB cells. The experiments were performed in triplicate and actin was used as a control

HOXA11-AS regulates epithelial to mesenchymal transition of skin cancer cells

The transwell assays showed that HOXA11-AS knockdown leads to a considerable decrease in the migration and invasion of A375 and HMCB cells (Fig. 6a, b).The protein expression levels of marker proteins of epithelial and mesenchymal cellular states were inspected through their blotting study. The expression of E-cadherin expressed was increased considerably while as the expression of N-cadherin decreased under the HOXA11-AS knockdown in both A375 and HMCB skin cancer cells (Fig. 6c). Thus, the results indicate that epithelial to mesenchymal transition in skin cancer cells is regulated by HOXA11-AS and its downregulation might actively tend to revert the epithelial to mesenchymal transition of skin cancer cells further advocating the therapeutic applicability of lncRNA HOXA11-AS against the human skin cancer.

Fig. 6.

Fig. 6

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HOXA11-AS silencing tend to revert the epithelial to mesenchymal transition in skin cancer cells, in vitro. a Transwell assay showing cell migration of si-NC and si-HOXA11-AS transfected A375 and HMCB cells b Transwell assay showing cell invasion assays of si-NC and si-HOXA11-AS transfected A375 and HMCB cells. c Western blotting of molecular markers of epithelial and mesenchymal cellular states of A375 and HMCB skin cancer cells transfected with si-HOXA11-AS or its negative control, si-NC. The experiments were performed in triplicate and actin was used as a control

Discussion

With the advent and subsequent advancements in the second-generation sequencing technologies, it became potentially possible to widen the exploration of regulatory mechanics of human cancers (Wong et al. 2011). Research studies indicated that the altered expression patterns of long-coding RNAs (lncRNA) surface the development and progression of human cancers (Zhang et al. 2016; Lin and Yang 2018). lncRNAs were found to regulate the critical steps of growth and proliferation of cancer cells (Yang et al. 2014). The malignant human skin cancer or melanoma has exceptionally low 5 year survival rate owing to extremely poor prognosis of the malignancy at earlier stages and higher metastatic potential of skin cancer cells (Schadendorf et al. 2018). Research findings have suggested the prognostic utility and therapeutic applicability of lncRNAs against the skin cancer (Aftab et al. 2014; Chen et al. 2016). The usefulness of lncRNAs as valuable and potential biomarkers in cancer diagnosis lies in the fact that their accessibility and detection is comparatively far easier to made using even the samples of urine and blood as the lncRNAs are secreted outside the cells in exosomes in very high concentrations (Gezer et al. 2014). Apart from that, it has been reported that the malignant cells show remarkably higher exosome secretions (Kharaziha et al. 2012). The lncRNAs secreted in exosomes might be utilized as fingerprints to specifically identify the malignant cells which will also aid in devising the selective targeting of such a cellular population (Ma et al. 2017). The gastric and pancreatic cancers have already been demonstrated for their identification using exosome-based diagnostic approach (Li et al. 2015; Işın et al. 2015). Although, lncRNAs have not been utilized in clinical setting in diagnosing the skin cancer, nevertheless there are several lncRNAs which are currently being tested for their applicability as biomarkers in skin cancer (Ma et al. 2017). In an investigation on diagnostic and therapeutic value of lncRNAs, we here found that HOXA11-AS was significantly up-regulated in skin cancer tissues and cancer cell lines. These inferences have been made regarding the expression analysis of HOXA11-AS for other cancer types also from the previous studies (Liu et al. 2017). HOXA11-AS, belonging to homeobox gene cluster, has been reported to be closely associated with the malignancy of the human tumors (Cui et al. 2017; Su and Hu 2017). In a previous study, the HOXA11-AS gene was shown to regulate the breast cancer growth and its knockdown was found to restrict the in vitro proliferation and viability of breast cancer cells (Li et al. 2017). Similar regulatory role was deduced for HOXA11-AS in human skin cancer from the present study. The decline in the proliferation of skin cancer cells was confirmed to be caused through the mitotic cell cycle arrest of skin cancer cells and the skin cancer cells were restricted for their mitotic entry. The higher rates of cell division of cancer cells result from the activation of several signaling pathways (Jin et al. 2012). The enhancement of the PI3K/AKT/mTOR signaling pathway is among many vital clues which are associated with the growth and progression of human cancers (Qiao et al. 2018). The phosphorylation acts as a crucial switch in activation of signaling pathways including the PI3K/AKT/mTOR signaling pathway (Kumar et al. 2005). The results of the present study revealed that HOXA11-AS regulates the phosphorylation of PI3K/AKT/mTOR pathway components and thus might act as an important drug target to indirectly target the signaling cascade, which researchers have advocated as an alternative therapeutic approach against the human cancers. Besides, the current study suggests that HOXA11-AS regulates the epithelial to mesenchymal transition of skin cancer cells. The epithelial to mesenchymal transition plays a fundamental role in invasion and metastasis of tumor cells (Kumar et al. 2005). At the molecular level, the transition leads to the loss of cell integrity and adhesion to the basement membrane enhancing the motility of tumor cells (Wittekind and Neid 2005). lncRNAs including HOXA11-AS have been reported to play critical roles in the molecular events of epithelial to mesenchymal transition (Li et al. 2017). Here in our study, we deduced that HOXA11-AS knockdown reduced the expression of mesenchymal marker proteins along with enhancing the protein expression of E-cadherin, an important biomarker of epithelial cellular state (Cates et al. 2008). Thus, HOXA11-AS might be having a regulatory role in controlling epithelial to mesenchymal transition of skin cancer cells. Summing up, the study suggests HOXA11-AS as an important prognostic and therapeutic biomolecule against the human skin cancer. The results of the current study are suggestive of utility of the HOXA11-AS gene to serve as vital prognostic molecule in skin cancer together with other lncRNAs reported earlier (Zhang et al. 2017).

Taken together, the results further support the potential of HOXA11-AS to act as therapeutic drug target in future to be employed against the human skin cancer. As the knockdown of HOXA11-AS inhibited the proliferation, migration and invasion of the skin cancer cells, it is obvious that in future the drug molecules specifically targeting HOXA11-AS might emerge as crucial alternative lead molecules against the human skin cancer. Therefore, the present study might be seen with a promise to pave way for many future research explorations in this direction. However, further in vivo studies are required to establish HOXA11-AS as the therapeutic target for the management of skin cancer.

Acknowledgements

The authors acknowledge the Cangzhou Central Hospital, Cangzhou, Hebei, China, 061001 for providing lab facilities.

Authors’ contributions

Conceptualization: JW and XL; methodology: JW and XL; formal analysis and investigation: JW, XL, HL and XiL; writing—original draft preparation: JW and XL; writing—review and editing critically for important intellectual content: JW and HL; supervision: JW.

Funding

This research did not obtain any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

All the authors declare that he has no conflict of interest.

Ethical approval

The study was approved by research ethics committee of Cangzhou Central Hospital, Cangzhou, Hebei, China on 6th of June, 2019 China with approval number CCH/0331/2019.

References

  1. Aftab MN, Dinger ME, Perera RJ. The role of microRNAs and long non-coding RNAs in the pathology, diagnosis, and management of melanoma. Arch Biochem Biophys. 2014;563:60–70. doi: 10.1016/j.abb.2014.07.022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Apalla Z, Lallas A, Sotiriou E, Lazaridou E, Ioannides D. Epidemiological trends in skin cancer. Dermatol Pract Concept. 2017;7(2):1–6. doi: 10.5826/dpc.0702a01. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Cates JM, Dupont WD, Barnes JW, Edmunds HS, Fasig JH, Olson SJ, Black CC. Markers of epithelial-mesenchymal transition and epithelial differentiation in sarcomatoid carcinoma: utility in the differential diagnosis with sarcoma. Appl Immunohistochem Mol Morphol. 2008;16(3):251–262. doi: 10.1097/PAI.0b013e318156e9b4. [DOI] [PubMed] [Google Scholar]
  4. Chen L, Yang H, Xiao Y, Tang X, Li Y, Han Q, Fu J, Yang Y, Zhu Y. LncRNA GAS5 is a critical regulator of metastasis phenotype of melanoma cells and inhibits tumor growth in vivo. Onco Targets Ther. 2016;9:4075–4087. doi: 10.2147/OTT.S98203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Cui M, Wang J, Li Q, Zhang J, Jia J, Zhan X. Long non-coding RNA HOXA11-AS functions as a competing endogenous RNA to regulate ROCK1 expression by sponging miR-124-3p in osteosarcoma. Biomed Pharmacother. 2017;92:437–444. doi: 10.1016/j.biopha.2017.05.081. [DOI] [PubMed] [Google Scholar]
  6. Di Gesualdo F, Capaccioli S, Lulli M. A pathophysiological view of the long non-coding RNA world. Oncotarget. 2014;5(22):10976–10996. doi: 10.18632/oncotarget.2770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dimitriou F, Krattinger R, Ramelyte E, Barysch MJ, Micaletto S, Dummer R, Goldinger SM. The world of melanoma: epidemiologic, genetic, and anatomic differences of melanoma across the globe. Curr Oncol Rep. 2018;20(11):87–91. doi: 10.1007/s11912-018-0732-8. [DOI] [PubMed] [Google Scholar]
  8. Gezer U, Özgür E, Cetinkaya M, Isin M, Dalay N. Long non-coding RNAs with low expression levels in cells are enriched in secreted exosomes. Cell Biol Inter. 2014;38(9):1076–1079. doi: 10.1002/cbin.10301. [DOI] [PubMed] [Google Scholar]
  9. Hauptman N, Glavac D. MicroRNAs and long non-coding RNAs: prospects in diagnostics and therapy of cancer. Radiol Oncol. 2013;47(4):311–318. doi: 10.2478/raon-2013-0062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Işın M, Uysaler E, Özgür E, Köseoğlu H, Şanlı Ö, Yücel ÖB, Gezer U, Dalay N. Exosomal lncRNA-p21 levels may help to distinguish prostate cancer from benign disease. Front Genet. 2015;6:168–171. doi: 10.3389/fgene.2015.00168. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jin YJ, Lee JH, Kim YM, Oh GT, Lee H. Macrophage inhibitory cytokine-1 stimulates proliferation of human umbilical vein endothelial cells by up-regulating cyclins D1 and E through the PI3K/Akt-, ERK-, and JNK-dependent AP-1 and E2F activation signaling pathways. Cell Signal. 2012;24(8):1485–1495. doi: 10.1016/j.cellsig.2012.03.014. [DOI] [PubMed] [Google Scholar]
  12. Kharaziha P, Ceder S, Li Q, Panaretakis T. (2012) Tumor cell-derived exosomes: a message in a bottle. Biochim Biophys Acta Rev Cancer. 2012;1826(1):103–111. doi: 10.1016/j.bbcan.2012.03.006. [DOI] [PubMed] [Google Scholar]
  13. Kumar V, Zhang MX, Swank MW, Kunz J, Wu GY. Regulation of dendritic morphogenesis by Ras–PI3K–Akt–mTOR and Ras–MAPK signaling pathways. J Neurosci. 2005;25(49):11288–11299. doi: 10.1523/JNEUROSCI.2284-05.2005. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Leiter U, Keim U, Eigentler T, Katalinic A, Holleczek B, Martus P, Garbe C. Incidence, mortality, and trends of nonmelanoma skin cancer in Germany. J Invest Dermatol. 2017;137(9):1860–1867. doi: 10.1016/j.jid.2017.04.020. [DOI] [PubMed] [Google Scholar]
  15. Li Q, Shao Y, Zhang X, Zheng T, Miao M, Qin L, Wang B, Ye G, Xiao B, Guo J. Plasma long noncoding RNA protected by exosomes as a potential stable biomarker for gastric cancer. Tumor Biol. 2015;36(3):2007–2012. doi: 10.1007/s13277-014-2807-y. [DOI] [PubMed] [Google Scholar]
  16. Li W, Jia G, Qu Y, Du Q, Liu B, Liu B. Long non-coding RNA (LncRNA) HOXA11-AS promotes breast cancer invasion and metastasis by regulating epithelial-mesenchymal transition. Med Sci Monitor. 2017;23:3393–3396. doi: 10.12659/MSM.904892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lin C, Yang L. Long noncoding RNA in cancer: wiring signaling circuitry. Trends Cell Biol. 2018;28(4):287–301. doi: 10.1016/j.tcb.2017.11.008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Liu Z, Chen Z, Fan R, Jiang B, Chen X, Chen Q, Nie F, Lu K, Sun M. Over-expressed long noncoding RNA HOXA11-AS promotes cell cycle progression and metastasis in gastric cancer. Mol Cancer. 2017;16(1):82–85. doi: 10.1186/s12943-017-0651-6. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lu S, Jiang X, Su Z, Cui Z, Fu W, Tai S. The role of the long non-coding RNA HOXA11-AS in promoting proliferation and metastasis of malignant tumors. Cell Biol Intr. 2018;42(12):1596–15601. doi: 10.1002/cbin.11045. [DOI] [PubMed] [Google Scholar]
  20. Ma X, He Z, Li L, Yang D. Liu G (2017) Expression profiles analysis of long non-coding RNAs identified novel lncRNA biomarkers with predictive value in outcome of cutaneous melanoma. Oncotarget. 2017;8(44):77761–77770. doi: 10.18632/oncotarget.20780. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Qiao J, Wang WJ, Zhang Y. Aclidinium inhibits proliferation and metastasis of ovarian cancer SKOV3 cells via downregulating PI3K/AKT/mTOR signaling pathway. Oncol Lett. 2018;16(5):6417–6422. doi: 10.3892/ol.2018.9460. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ribero S, Glass D, Bataille V. Genetic epidemiology of melanoma. Eur J Dermatol. 2016;26(4):335–359. doi: 10.1684/ejd.2016.2787. [DOI] [PubMed] [Google Scholar]
  23. Richards EJ, Permuth-Wey J, Li Y, Chen YA, Coppola D, Reid BM, Lin HY, Teer JK, Berchuck A, Birrer MJ, Lawrenson K. A functional variant in HOXA11-AS, a novel long non-coding RNA, inhibits the oncogenic phenotype of epithelial ovarian cancer. Oncotarget. 2015;6(33):34745–34757. doi: 10.18632/oncotarget.5784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schadendorf D, van Akkooi AC, Berking C, Griewank KG, Gutzmer R, Hauschild A, Stang A, Roesch A, Ugurel S. Melanoma Lancet. 2018;392(10151):971–984. doi: 10.1016/S0140-6736(18)31559-9. [DOI] [PubMed] [Google Scholar]
  25. Su JC, Hu XF. Long non-coding RNA HOXA11-AS promotes cell proliferation and metastasis in human breast cancer. Mol Med Rep. 2017;16(4):4887–4894. doi: 10.3892/mmr.2017.7163. [DOI] [PubMed] [Google Scholar]
  26. Sun M, Liu XH, Lu KH, Nie FQ, Xia R, Kong R, Yang JS, Xu TP, Liu YW, Zou YF, Lu BB. EZH2-mediated epigenetic suppression of long noncoding RNA SPRY4-IT1 promote s NSCLC cell proliferation and metastasis by affecting the epithelial–mesenchymal transition. Cell Death Dis. 2014;5(6):e1298. doi: 10.1038/cddis.2014.256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wei C, Zhao L, Liang H, Zhen Y, Han L. Recent advances in unraveling the molecular mechanisms and functions of HOXA11-AS in human cancers and other diseases. Oncol Rep. 2020;43(6):1737–1754. doi: 10.3892/or.2020.7552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Whiteman DC, Green AC, Olsen CM. The growing burden of invasive melanoma: projections of incidence rates and numbers of new cases in six susceptible populations through 2031. J Investig Dermatol. 2016;136(6):1161–1171. doi: 10.1016/j.jid.2016.01.035. [DOI] [PubMed] [Google Scholar]
  29. Wittekind C, Neid M. Cancer invasion and metastasis. Oncology. 2005;69(Suppl. 1):14–16. doi: 10.1159/000086626. [DOI] [PubMed] [Google Scholar]
  30. Wong KM, Hudson TJ, McPherson JD. Unraveling the genetics of cancer: genome sequencing and beyond. Annu Rev Genomics Hum Genet. 2011;12:407–430. doi: 10.1146/annurev-genom-082509-141532. [DOI] [PubMed] [Google Scholar]
  31. Xu Q, Deng F, Qin Y, Zhao Z, Wu Z, Xing Z, Ji A, Wang QJ. Long non-coding RNA regulation of epithelial–mesenchymal transition in cancer metastasis. Cell Death Dis. 2016;7(6):e2254. doi: 10.1038/cddis.2016.149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yang G, Lu X. Yuan L (2014) LncRNA: a link between RNA and cancer. Bioochim Biophys Acta Gene Regu Mech. 1839;11:1097–1109. doi: 10.1016/j.bbagrm.2014.08.012. [DOI] [PubMed] [Google Scholar]
  33. Zhang R, Xia LQ, Lu WW, Zhang J, Zhu JS. LncRNAs and cancer. Oncol Lett. 2016;12(2):1233–1239. doi: 10.3892/ol.2016.4770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zhang Y, Chen WJ, Gan TQ, Zhang XL, Xie ZC, Ye ZH, Deng Y, Wang ZF, Cai KT, Li SK, Luo DZ. Clinical significance and effect of lncRNA HOXA11-AS in NSCLC: a study based on bioinformatics, in vitro and in vivo verification. Sci Rep. 2017;7(1):1–8. doi: 10.1038/s41598-016-0028-x. [DOI] [PMC free article] [PubMed] [Google Scholar]

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