Skip to main content
NCBI home page
The Journal of International Advanced Otology logoLink to The Journal of International Advanced Otology
. 2023 May 1;19(3):212–216. doi: 10.5152/iao.2023.22863

Expression and Correlation Research of MicroRNA-10a-5p and PIK3CA in Middle Ear Cholesteatoma

Jing Yang 1,2, Wei Yan 1,3, Susu Tang 1, Zhikun Huang 1, Mingyuan Ye 1, Zheng Lu 1, Qianxu Liu 1,✉
PMCID: PMC10331635  PMID: 37272638

Abstract

Background:

This study aimed to examine the roles of miR-10a-5p and phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α in the pathogenesis of middle ear cholesteatoma.

Methods:

We enrolled 27 patients with middle ear cholesteatoma and collected samples of intraoperative cholesteatoma and normal posterior ear skin tissues. The mRNA expression levels of miR-10a-5p and PIK3CA were detected using real-time quantitative polymerase chain reaction. PIK3CA protein expression was measured by immunohistochemistry and western blotting.

Results:

Middle ear cholesteatoma tissues showed significantly lower miR-10a-5p expression levels and significantly higher PIK3CA expression levels than normal posterior ear skin tissues (both P < .05). Furthermore, the miR-10a-5p and PIK3CA expression levels were significantly negatively correlated in middle ear cholesteatoma tissues (r = −0.926, P < .001).

Conclusion:

Low miR-10a-5p expression levels in middle ear cholesteatoma tissues may inhibit the growth and proliferation of cholesteatoma, whereas high PIK3CA expression level may promote its growth and proliferation. In addition, miR-10a-5p may affect the proliferation and differentiation of cholesteatoma by negatively regulating its target gene, PIK3CA.

Keywords: Cholesteatoma, gene expression, microRNAs, middle ear

Introduction

Middle ear cholesteatoma, a benign cystic lesion, is a frequently occurring condition in otorhinolaryngology.1 It is characterized by abnormally growing keratinized squamous epithelium within the temporal bone, which can gradually destroy the surrounding bone. Middle ear cholesteatoma can be associated with intracranial and extracranial complications, including hearing loss, vertigo, facial paralysis, and brain abscess.2,3 Surgical resection is the only effective method for treating middle ear cholesteatoma. However, surgery is associated with certain risks, including those of facial paralysis, vertigo, and recurrence.4 Therefore, it is necessary to explore nonsurgical treatment options for cholesteatoma.5

Recently, more studies have investigated the role of microRNA (miRNA) in the occurrence and development of middle ear cholesteatoma. Xie et al6 used miRNA microarray technology to study expression profiles of ear cholesteatoma and normal skin tissues and found that the expression of miR-10a-5p was significantly different between these tissues. They also revealed that phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) might be the most important target gene of miR-10a-5p. PIK3CA, a catalytic subunit of PI3K, is a key activation molecule in the EGFR-PI3K-Akt-CyclinD1 signaling pathway, which is important in regulating cholesteatoma.7 The involvement of PIK3CA mutations has been reported in the progression of breast cancer,8 kidney cancer,9 and squamous cell carcinoma in the head and neck,10 among others.

Middle ear cholesteatoma is benign yet exhibits patterns of proliferation, invasion, and migration, similar to those observed in malignant tumors.11 We speculate that miR-10a-5p and PIK3CA play important roles in the development and growth of cholesteatoma. Currently, to the best of our knowledge, there is no literature regarding the mechanisms of action of miR-10a-5p and PIK3CA in middle ear cholesteatoma. Therefore, we chose miR-10a-5p as the research object and aimed to examine and compare the expression of miR-10a-5p and PIK3CA in cholesteatoma and normal skin tissues from behind the ear; to explore the effects of their expression on cholesteatoma; and to provide insights for nonsurgical treatment options of middle ear cholesteatoma.

Methods

Participants

In this study, we enrolled 27 patients [18 female and 9 male participants aged 5–76 years (mean ± standard deviation, 40.4 ± 16.6 years)] with acquired secondary middle ear cholesteatoma diagnosed at Zhuhai People’s Hospital (Zhuhai Hospital Affiliated with Jinan University) from June 2020 to January 2021. The participants were receiving their initial surgical treatment during the study period. This study was conducted according to tenets of the Helsinki Declaration of 1975, as revised in 2008, and was approved by the Ethics Committee of Zhuhai People’s Hospital (No. 69 of 2020). All 27 patients or their guardians provided written informed consent. The study adhered to the TREND guidelines for nonrandomized ­public behavior.

Research Methods

Reagents

The total RNA extraction reagent (TRIzol® Reagent) and RT Reagent Kit were purchased from Takara (Kyoto, Japan); the Bulge-Loop™ miRNA qRT-PCR Starter Kit from Guangzhou RiboBio Co., Ltd. (Guangzhou, China); PIK3CA primary antibody from Abcam Trading Co., Ltd. (Shanghai, China); general SP kit and diaminobenzidine (DAB) color development kits from Beijing Zhongshan Jinqiao Biotechnology Co., Ltd. (Beijing, China); WB Super RIPA Lysis Buffer, bicinchoninic acid (BCA) protein quantification kits, and Benzonase® Nuclease from HaiGene (Harbin, China); and gel preparation kit, western primary and secondary antibody dilutions, and sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein loading buffer from Beyotime Biotechnology Co., Ltd. (Shanghai, China).

Total RNA Extraction, Reverse Transcription, and Real-Time Quantitative Polymerase Chain Reaction

Total RNA was extracted from the matrix of middle ear cholesteatoma and retroauricular normal skin tissues according to the Trizol test method.12 RNA content was measured using UV spectrophotometry, and its integrity was evaluated using agarose gel electrophoresis. The extracted total RNA was reverse-transcribed into cDNA and amplified by polymerase chain reaction (PCR), according to the instructions of the real-time quantitative reverse transcription PCR (qRT-PCR) detection kit. We subsequently used the Bulge-Loop™ miRNA qRT-PCR Starter Kit to perform PCR. U6 was used as the internal reference gene for detecting miR-10a-5p, and single well system for each qRT-PCR reaction was 20 µL. We used GAPDH as the internal reference gene for detecting PIK3CA. The primer sequences were as follows: hsa-miR-10a-5p-RT, 5¢-GTCGTATCCAGTGCGTGTCGTGGAGTCGGCAATTGCACTGGATACGACCACAAAT-3¢; hsa-miR-10a-5p F, 5¢-GGTACCCTGTAGATCCGAATT-3¢; hsa-miR-10a-5p R, 5¢-CAGTGCGTGCGTGGAGT-3¢; PIK3CA F, 5¢-ACTATCCCCGAAATTCTACCCAA-3¢; and PIK3CA R, 5¢-AGGCCAATCTTTTACCAAGCAA-3¢. We repeated these reactions 3 times and calculated the test result using the 2-∆∆CT value.

Immunohistochemistry

Fresh cholesteatoma and skin samples collected during surgery were dehydrated with alcohol, embedded in paraffin, and serially sectioned at a thickness of 3 μm. The sections were stained with DAB and hematoxylin and then blocked. The samples were incubated with 100 μL of PIK3CA primary antibody diluted at 1 : 50 ratio at 4°C and then incubated with 100 μL of biotinylated goat antimouse/rabbit immunoglobulin G polymer (secondary antibody) at 20-25°C. We used known positive sections as positive controls and normal external auditory canal skin samples as negative controls. The primary antibody was replaced with Tris-buffered saline containing 0.020% Triton X-100 to exclude nonspecific staining.

Western Blot Detection

We powdered 20 mg of tissues and added 200 μL of WB Super RIPA Lysis Buffer to extract the total protein in each group. The BCA method was used to detect protein concentration.13 Samples were denatured at 100°C for 10 minutes, followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and were subsequently transferred onto a polyvinylidene difluoride membrane. The polyvinylidene difluoride membrane was blocked in 5% skim milk for 2 hours. We added a 1 : 500-fold dilution of the primary antibody and incubated the samples at 4°C overnight. The secondary antibody was incubated for 2 hours at 20-25°C, the film was washed, and the color was allowed to develop by chemiluminescence. Grayscale analysis was performed using ImageJ software (National Institutes of Health, Bethesda, Md, USA). The ratio of the grayscale value of the target band (PIK3CA) to that of the internal reference GAPDH band denoted the relative protein expression content.

Statistical Analyses

For data analysis, Statistical Package for Social Sciences version 26.0 (IBM SPSS Corp., Armonk, NY, USA) and GraphPad Prism version 7.0 (GraphPad Software, San Diego, CA, USA) software packages were used. Normally distributed data are presented as means and standard deviations. Unpaired t-tests were used to compare 2 independent samples. We used Pearson correlation analysis to compare the relative expression of 2 genes. P values of <.05 were considered statistically significant.

Results

Bioinformatics Methods for Predicting the Target Genes of miR-10a-5p

Target gene prediction results showed that the 3′ untranslated regions of PIK3CA had conserved binding sites to miR-10a-5p, suggesting that PIK3CA might be a downstream target gene of miR-10a-5p (Supplementary Figures 1 and 2).

Results of qRT-PCR Analysis for miR-10a-5p and PIK3CA mRNA

This experiment was strictly included in the qRT-PCR analysis standard following the Starter Kit instructions. After obtaining samples with total RNA purity (A260/A280) ranging from 1.8 to 2.0, 17 middle ear cholesteatoma tissue samples and 13 normal posterior ear skin tissue samples were analyzed using qRT-PCR. The results displayed the typical S-type fluorescence quantitative amplification curve, and the dissolution curve was unimodal. No other abnormal waveforms were noted, and the shape of the wave crest was sharp. These results indicated that the product was well amplified; moreover, no nonspecific amplification was observed (Supplementary Figures 3 and 4).

Both miR-10a-5p and PIK3CA mRNA were expressed in the cholesteatoma and normal posterior ear skin tissues. The miR-10a-5p expression level in cholesteatoma was significantly lower than that in the posterior skin (t = 2.208, P = .036) (Table 1, Supplementary Figure 5). In contrast, the PIK3CA mRNA expression level was significantly higher in the cholesteatoma than in the posterior skin (t = 3.951, P < .001) (Table 2, Supplementary Figure 6).

Table 1.

miR-10a-5p Expression in Patients with Middle Ear Cholesteatoma and Normal Skin Tissues

Group n miR-10a-5p t P
Middle ear cholesteatoma tissue 17 0.99 ± 0.03 2.208 .036
Normal posterior ear skin tissue 13 1.61 ± 0.18
Open in a new tab

Data are presented as mean±standard deviation

Table 2.

Relative Expression of PIK3CA mRNA in Patients with Middle Ear Cholesteatoma and Normal Skin Tissues

Group n PIK3CA t P
Middle ear cholesteatoma tissue 17 2.06±0.23 3.951 <.001
Normal posterior ear skin tissue 13 1.02±0.05
Open in a new tab

Data are presented as mean ± standard deviation

PIK3CA, phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α.

Immunohistochemical Analysis of PIK3CA

In middle ear cholesteatoma, PIK3CA was primarily located in the cytoplasm and cell membrane; its expression may have been accompanied by partial nuclear staining. It was expressed from the basal layer to the upper basal layer. The staining degree was mostly moderate to strong, and the positive expression of PIK3CA was indicated by yellow or brownish-yellow findings (Figure 1). Conversely, in normal skin tissues, PIK3CA was primarily located in the nucleus and cell membrane. Its expression might be accompanied by partial cytoplasmic staining, which was characterized by the staining of cells in the basal layer and a few suprabasal layers. In contrast to the amount of staining observed in cholesteatoma tissues, the amount of staining noted in normal skin tissues was mainly weakly positive. The positive expression of PIK3CA was indicated by yellow or brownish-yellow findings (Figure 2).

Figure 1.

Figure 1.

Open in a new tab

Expression of phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) in middle ear cholesteatoma (A: ×200; B: ×400).

Figure 2.

Figure 2.

Open in a new tab

Expression of phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) in normal posterior ear skin (A: ×200; B: ×400).

Western Blot Analysis of PIK3CA

Protein blots of PIK3CA were present in both the experimental and control specimens, with significantly higher PIK3CA expression in cholesteatoma tissue samples than in normal posterior ear skin tissue samples (P < .05) (Figure 3). These results were also consistent with the qRT-PCR detection results.

Figure 3.

Figure 3.

Open in a new tab

Expression of phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) protein in middle ear cholesteatoma and retroauricular normal skin tissues (*P < 0.0).

Pearson Correlation Analysis of miR-10a-5p and PIK3CA mRNA Expression in Middle Ear Cholesteatoma

Pearson correlation analysis was used to determine the correlation between miR-10a-5p and PIK3CA mRNA expression in middle ear cholesteatoma, and the results indicated a significant negative correlation (r = −0.926, P < .001) (Figure 4).

Figure 4.

Figure 4.

Open in a new tab

Correlation analysis of the expression of miR-10a-5p and phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) mRNA in middle ear cholesteatoma.

Discussion

Middle ear cholesteatoma may be harmful and may require surgical treatment, which has associated risks. The development of a new treatment for middle ear cholesteatoma would have substantial clinical benefits. The current research focus is on gene therapy.

Researchers have conducted several studies on the role of miRNA in tumors. Li et al14 showed that miRNA-21 was significantly upregulated in cancer cells from various sources and that miRNA-21 regulated the initiation, transformation, invasion, and metastasis of cancer. Yoshikawa et al15 intraperitoneally injected synthetic miR-214 into mice with canine angiosarcoma and observed that it produced antitumor effects by inducing apoptosis and inhibiting cell proliferation. In addition, miRNA expression has been shown to affect the occurrence, development, and prognosis of various malignancies, including lung cancer,16 prostate cancer,17 and liver cancer.18 Since the critical role of miRNAs in cancer has been demonstrated, gene therapy based on miRNA has attracted considerable attention.19,20

Since Friedland et al21 first reported the expression of miRNA-21 and its downstream target genes PTEN and PDCD4 in middle ear cholesteatoma and normal skin tissues in 2009, researchers have reported that miRNA is involved in regulating the proliferation, apoptosis, invasion, bone destruction, angiogenesis, and other pathophysiological processes of cholesteatoma cells.22-24 Moreover, miR-10a-5p belongs to the miR-10 family and is downregulated in various malignant tumors, such as cervical cancer,25 renal cell carcinoma,26 and liver cancer.27

Double luciferase reporter gene testing in breast cancer MCF-7 cells by Zhang et al28 showed that miR-10a-5p could directly target the 3’ untranslated region of PIK3CA. PIK3CA mutations play an important part in cell proliferation, survival, and tumorigenesis by activating the PI3K-Akt signaling pathway.29,30 Liu et al7 confirmed that the EGFR-PI3K-Akt-cyclin D1 signaling pathway is active in cholesteatoma and may play a vital role in cholesteatoma epithelial hyperproliferation.

The qRT-PCR findings in the present study indicate that the miR-10a-5p expression level in middle ear cholesteatoma tissues was lower than that in normal skin tissues. Our results are consistent with those of Xie et al.6 who conducted microarray analysis and qRT-PCR verification. Vaher et al31 showed that miR-10a-5p could inhibit keratinocyte proliferation and regulate cell cycle. Therefore, the results of this experiment are consistent with those on miR-10a-5p inhibition in the growth and proliferation of middle ear cholesteatoma.

In this study, the expression level of PIK3CA mRNA was significantly higher in middle ear cholesteatoma tissues than that in normal skin tissues according to qRT-PCR and Western blot findings. Immunohistochemistry showed that PIK3CA staining was mostly moderate to strongly positive in middle ear cholesteatoma but was mostly weakly positive in normal skin tissues sampled from behind the ear. While we did not retrieve reports regarding the expression of PIK3CA in cholesteatoma, we found that PIK3CA was highly expressed in various malignancies, such as gastric cancer,32 breast cancer,33 and cervical cancer,34 and promoted the growth of these tumors. We hypothesized that PIK3CA also promotes the proliferation of cholesteatoma based on the many malignant characteristics of cholesteatoma. The high expression of PIK3CA in cholesteatoma detected in this study is in line with the logic that PIK3CA promotes the proliferation of cholesteatoma.

Our results demonstrated that miR-10a-5p expression was negatively correlated with PIK3CA expression in patients with middle ear cholesteatoma, suggesting that downregulated miR-10a-5p in middle ear cholesteatoma may negatively regulate the function of its target gene PIK3CA and further activate the PI3K-AKT signaling pathway to accelerate the proliferation and differentiation of cholesteatoma cells. Zhang et al28 reported that the expression of PIK3CA could be downregulated by increasing the expression of miR-10a-5p in breast cancer cells. Downregulation inhibits activation of the PI3K-AKT signaling pathway, ultimately suppressing cell proliferation and promoting cell apoptosis.

This study has some limitations. We only used a small number of clinical samples, and we did not examine the regulatory mechanisms. Future studies should replicate these experiments with a larger sample size. In addition, further studies should prove whether miR-10a-5p can reduce the activation of PIK3CA and can inhibit the proliferation of cholesteatoma and evaluate whether the underlying mechanism of miR-10a-5p and PIK3CA regulates the proliferation and differentiation of middle ear cholesteatoma.

Conclusion

This study confirmed that low miR-10a-5p expression and high PIK3CA expression in middle ear cholesteatoma were negatively correlated. miR-10a-5p may affect the proliferation and differentiation of cholesteatoma by negatively regulating PIK3CA. Our study provides a novel approach to nonsurgical treatment options for middle ear cholesteatoma.

Supplementary Figure 1.

Supplementary Figure 1.

Open in a new tab

The TargetScan software predicts phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) as a target gene for miR-10a-5p.

Supplementary Figure 2.

Supplementary Figure 2.

Open in a new tab

The miRBase software predicts phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) as a target gene for miR-10a-5p.

Supplementary Figure 3.

Supplementary Figure 3.

Open in a new tab

Amplification and dissolution curves of miR-10a-5p.

Supplementary Figure 4.

Supplementary Figure 4.

Open in a new tab

Amplification and dissolution curves of phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) mRNA.

Supplementary Figure 5.

Supplementary Figure 5.

Open in a new tab

Expression of miR-10a-5p in middle ear cholesteatoma and normal posterior ear skin tissues (*P < .05).

Supplementary Figure 6.

Supplementary Figure 6.

Open in a new tab

Expression of phosphatidylinositol-4,5-bisphosphonate 3-kinase catalytic subunit α (PIK3CA) mRNA in middle ear cholesteatoma and normal posterior ear skin tissues (**P < .001).

Footnotes

Ethics Committee Approval: Ethical committee approval was received from the Ethics Committee of Zhuhai People’s Hospital (No. 69 of 2020).

Informed Consent: Written informed consent was obtained from all patients or their guardians. .

Peer-review: Externally peer-reviewed.

Author Contributions: Concept – Q.L., J.Y.; Design – J.Y., S.T.; Supervision – Q.L., W.Y.; Funding – W.Y., S.T.; Materials – S.T., Z.H.; Data Collection and/or Processing – Z.H., M.Y.; Analysis and/or Interpretation – M.Y., Z.L.; Literature Review – J.Y., Z.L.; Writing – J.Y.; Critical Review –Q.L.

Acknowledgments: Shengni Hua helped in designing the study, and Zhao Liu contributed to the collection of cholesteatoma specimens from the Department of Otorhinolaryngology, Zhuhai People’s Hospital. We express our heartfelt gratitude for their help. We would also like to thank Editage (www.editage.cn) for English language editing.

Declaration of Interests: The authors declare that they have no competing interest.

Funding: This study was supported by the Medical Scientific Research Foundation of Zhuhai, Guangdong, China (Zhuhai Science and Technology Innovation Bureau, grant number ZH22036201210091PWC).

References

  • 1. Louw L. Acquired cholesteatoma pathogenesis: stepwise explanations. J Laryngol Otol. 2010;124(6):587 593. ( 10.1017/S0022215109992763) [DOI] [PubMed] [Google Scholar]
  • 2. Maniu A, Harabagiu O, Perde Schrepler M, Cătană A, Fănuţă B, Mogoantă CA. Molecular biology of cholesteatoma. Rom J Morphol Embryol. 2014;55(1):7 13. [PubMed] [Google Scholar]
  • 3. Prasad SC, Piras G, Piccirillo E, et al. Surgical strategy and facial nerve outcomes in petrous bone cholesteatoma. Audiol Neurootol. 2016;21(5):275 285. ( 10.1159/000448584) [DOI] [PubMed] [Google Scholar]
  • 4. Luntz M, Barzilai R. Middle ear cholesteatoma [in Hebrew]. Harefuah. 2021;160(5):316 322. [PubMed] [Google Scholar]
  • 5. Uzun T, Çaklı H, Coşan DT, et al. In vitro study on immune response modifiers as novel medical treatment options for cholesteatoma. Int J Pediatr Otorhinolaryngol. 2021;145:110743. ( 10.1016/j.ijporl.2021.110743) [DOI] [PubMed] [Google Scholar]
  • 6. Xie S, Liu X, Pan Z, et al. Microarray analysis of differentially expressed microRNAs in acquired middle ear cholesteatoma. Int J Med Sci. 2018;15(13):1547 1554. ( 10.7150/ijms.26329) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. Liu W, Ren H, Ren J, et al. The role of EGFR/PI3K/Akt/cyclinD1 signaling pathway in acquired middle ear cholesteatoma. Mediators Inflamm. 2013;2013:651207. ( 10.1155/2013/651207) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8. Dumbrava EE, Call SG, Huang HJ, et al. PIK3CA mutations in plasma circulating tumor DNA predict survival and treatment outcomes in patients with advanced cancers. ESMO Open. 2021;6(5):100230. ( 10.1016/j.esmoop.2021.100230) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9. Gasinska A, Jaszczynski J, Adamczyk A, et al. Biomarkers of epithelial-mesenchymal transition in localized, surgically treated clear-cell renal cell carcinoma. Folia Histochem Cytobiol. 2018;56(4):195 206. ( 10.5603/FHC.a2018.0023) [DOI] [PubMed] [Google Scholar]
  • 10. Jin N, Keam B, Cho J, et al. Therapeutic implications of activating noncanonical PIK3CA mutations in head and neck squamous cell carcinoma. J Clin Invest. 2021;131(22):e150335. ( 10.1172/JCI150335) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 11. Kuo CL. Etiopathogenesis of acquired cholesteatoma: prominent theories and recent advances in biomolecular research. Laryngoscope. 2015;125(1):234 240. ( 10.1002/lary.24890) [DOI] [PubMed] [Google Scholar]
  • 12. Brown RAM, Epis MR, Horsham JL, Kabir TD, Richardson KL, Leedman PJ. Total RNA extraction from tissues for microRNA and target gene expression analysis: not all kits are created equal. BMC Biotechnol. 2018;18(1):16. ( 10.1186/s12896-018-0421-6) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13. Komatsu S. Western blotting using PVDF membranes and its downstream applications. Methods Mol Biol. 2015;1312:227 236. ( 10.1007/978-1-4939-2694-7_24) [DOI] [PubMed] [Google Scholar]
  • 14. Li S, Liang Z, Xu L, Zou F. MicroRNA-21: a ubiquitously expressed pro-survival factor in cancer and other diseases. Mol Cell Biochem. 2012;360(1-2):147 158. ( 10.1007/s11010-011-1052-6) [DOI] [PubMed] [Google Scholar]
  • 15. Yoshikawa R, Maeda A, Ueno Y, et al. Intraperitoneal administration of synthetic microRNA-214 elicits tumor suppression in an intraperitoneal dissemination mouse model of canine hemangiosarcoma. Vet Res Commun. 2022;46(2):447 457. ( 10.1007/s11259-021-09869-1) [DOI] [PubMed] [Google Scholar]
  • 16. Zhao J, Yu H, Han T, Zhu X. Prognosis value of microRNA-3677-3p in lung adenocarcinoma and its regulatory effect on tumor progression. Cancer Manag Res. 2021;13:9261 9270. ( 10.2147/CMAR.S330357) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17. Olivan M, Garcia M, Suárez L, et al. Loss of microRNA-135b enhances bone metastasis in prostate cancer and predicts aggressiveness in human prostate samples. Cancers (Basel). 2021;13(24):6202. ( 10.3390/cancers13246202) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 18. Yugawa K, Yoshizumi T, Mano Y, et al. Cancer-associated fibroblasts promote hepatocellular carcinoma progression through downregulation of exosomal miR-150-3p. Eur J Surg Oncol. 2021;47(2):384 393. ( 10.1016/j.ejso.2020.08.002) [DOI] [PubMed] [Google Scholar]
  • 19. Wallace JA, O’Connell RM. MicroRNAs and acute myeloid leukemia: therapeutic implications and emerging concepts. Blood. 2017;130(11):1290 1301. ( 10.1182/blood-2016-10-697698) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. Trobaugh DW, Klimstra WB. MicroRNA regulation of RNA virus replication and pathogenesis. Trends Mol Med. 2017;23(1):80 93. ( 10.1016/j.molmed.2016.11.003) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. Friedland DR, Eernisse R, Erbe C, Gupta N, Cioffi JA. Cholesteatoma growth and proliferation: posttranscriptional regulation by microRNA-21. Otol Neurotol. 2009;30(7):998 1005. ( 10.1097/MAO.0b013e3181b4e91f) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22. Zhang W, Chen X, Qin Z. MicroRNA let-7a suppresses the growth and invasion of cholesteatoma keratinocytes. Mol Med Rep. 2015;11(3):2097 2103. ( 10.3892/mmr.2014.2971) [DOI] [PubMed] [Google Scholar]
  • 23. Li Y, Liang J, Hu J, Ren X, Sheng Y. Down-regulation of exosomal miR-106b-5p derived from cholesteatoma perimatrix fibroblasts promotes angiogenesis in endothelial cells by overexpression of angiopoietin 2. Cell Biol Int. 2018;42(10):1300 1310. ( 10.1002/cbin.11002) [DOI] [PubMed] [Google Scholar]
  • 24. Zang J, Hui L, Yang N, Yang B, Jiang X. Downregulation of MiR-203a disinhibits bmi1 and promotes growth and proliferation of keratinocytes in cholesteatoma. Int J Med Sci. 2018;15(5):447 455. ( 10.7150/ijms.22410) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 25. Zhai L, Li Y, Lan X, Ai L. MicroRNA-10a-5p suppresses cancer proliferation and division in human cervical cancer by targeting BDNF. Exp Ther Med. 2017;14(6):6147 6151. ( 10.3892/etm.2017.5312) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26. Arai T, Okato A, Kojima S, et al. Regulation of spindle and kinetochore-associated protein 1 by antitumor miR-10a-5p in renal cell carcinoma. Cancer Sci. 2017;108(10):2088 2101. ( 10.1111/cas.13331) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. Zheng X, Li JW, Liu YK, et al. microRNA-10a-5p overexpression suppresses malignancy of colon cancer by regulating human liver cancer fibroblasts. Neoplasma. 2021;68(6):1157 1168. ( 10.4149/neo_2021_210226N250) [DOI] [PubMed] [Google Scholar]
  • 28. Zhang J, Ren L, Yu M, et al. S-equol inhibits proliferation and promotes apoptosis of human breast cancer MCF-7 cells via regulating miR-10a-5p and PI3K/AKT pathway. Arch Biochem Biophys. 2019;672:108064. ( 10.1016/j.abb.2019.108064) [DOI] [PubMed] [Google Scholar]
  • 29. Mayer IA, Arteaga CL. The PI3K/AKT pathway as a target for cancer treatment. Annu Rev Med. 2016;67:11 28. ( 10.1146/annurev-med-062913-051343) [DOI] [PubMed] [Google Scholar]
  • 30. Trautmann M, Cyra M, Isfort I, et al. Phosphatidylinositol-3-kinase (PI3K)/Akt signaling is functionally essential in myxoid liposarcoma. Mol Cancer Ther. 2019;18(4):834 844. ( 10.1158/1535-7163.MCT-18-0763) [DOI] [PubMed] [Google Scholar]
  • 31. Vaher H, Runnel T, Urgard E, et al. miR-10a-5p is increased in atopic dermatitis and has capacity to inhibit keratinocyte proliferation. Allergy. 2019;74(11):2146 2156. ( 10.1111/all.13849) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32. Pai SM, Huang KH, Chen MH, et al. Cardia gastric cancer is associated with increased PIK3CA amplifications and HER2 expression than noncardia gastric cancer according to Lauren classification. Front Oncol. 2021;11:632609. ( 10.3389/fonc.2021.632609) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. Coussy F, El Botty R, Lavigne M, et al. Combination of PI3K and MEK inhibitors yields durable remission in PDX models of PIK3CA-mutated metaplastic breast cancers. J Hematol Oncol. 2020;13(1):13. ( 10.1186/s13045-020-0846-y) [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 34. Lachkar B, Minaguchi T, Akiyama A, et al. Prognostic significance of PIK3CA mutation in stage IIB to IVA cervical cancers treated by concurrent chemoradiotherapy with weekly cisplatin. Med (Baltim). 2018;97(31):e11392. ( 10.1097/MD.0000000000011392) [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of International Advanced Otology are provided here courtesy of The European Academy of Otology and Neurotology

RESOURCES