Abstract
Chemokines and their receptors have been proposed to play important roles in tumor progression and metastasis. To investigate their roles in the progression of primary and metastatic malignant liver tumors and their prognosis, we compared expression profiles of CXCL12/CXCR4, CCL20/CCR6, and CCL21/CCR7 in hepatocellular carcinoma (HCC) and colorectal liver metastases (CRLM). Immunohistochemistry was used to analyze the expression levels of the chemokine/chemokine receptor pairs in 29 HCC and 11 CRLM specimens and adjacent non-cancerous tissues, and correlations with clinicopathological variables and overall survival were determined. CCL20/CCR6 expression was higher in HCC than in adjacent non-cancerous tissues. High CCR6 expression in HCC was negatively associated with 5-year survival rate and was an independent prognostic factor for overall survival of HCC patients, whereas differences were not observed between CRLM and adjacent tissues. Furthermore, significantly higher expression of CCL21/CCR7 was found in CRLM than in HCC. In summary, the CCL20/CCR6 axis was elevated in HCC but not in CRLM, whereas the CCL21/CCR7 axis was elevated in CRLM but not in HCC.
Keywords: chemokines, chemokine receptors, colorectal liver metastases, hepatocellular carcinoma, prognosis
Introduction
As a family of small chemotactic cytokines, chemokines have traditionally been known as mediators of leukocyte trafficking and homing. However, accumulating evidence has demonstrated that chemokines and their receptors serve as key regulators of the tumor microenvironment by autocrine or paracrine mechanisms, which in turn promote tumor growth, cellular migration and invasion, angiogenesis, and distant spread of malignant tumors.1,2
Several studies have applied a variety of experimental methods such as quantitative real-time polymerase chain reaction (qRT-PCR), Western blot analysis, enzyme-linked immunosorbent assay (ELISA), and immunohistochemistry (IHC) to assess mRNA and protein expression profiles and intracellular and extracellular localization patterns of chemokines and their receptors in resected tissue samples of primary malignant hepatocellular carcinoma (HCC) and secondary malignant colorectal liver metastases (CRLM), which are found in up to 95% of patients with colorectal cancer (CRC) in the advanced disease stage.3–7 Various chemokine/chemokine receptor pairs can promote occurrence and development of HCC and/or CRLM, suggesting that these chemokine/chemokine receptor pairs may be used for prediction, prognostic evaluation, and differential diagnosis of HCC and CRLM. The chemokine/chemokine receptor pairs may be potential targets for novel treatment strategies because, for example, CCL20 and/or CCR6 expression has been reported to play an important role in carcinogenesis, progression, and intrahepatic metastasis of HCC8–14 and CRLM.8,15,16 A meta-analysis indicated that CXCR4 expression was associated with poor survival of HCC patients,17 and it has been reported to be a predictive factor for CRLM.18,19 The CCL21/CCR7 axis has been shown to play an important role in lymphoid organ development, tumor cell migration, and lymph node metastasis of several types of cancers, but studies on the relationship between this axis and the development of HCC and CRLM are rare.18,20–22 Furthermore, the analysis of RNA and protein expression levels of the chemokines CXCL12, CCL19, CCL20, and CCL21 and their receptors in HCC and CRLM demonstrated that CXCR4 expression levels can be used to differentiate between primary and secondary liver tumors, whereas CCL20 and its receptor CCR6 may promote tumor growth and may be used to differentiate between primary and secondary liver tumors.15
Expression of CXCL12/CXCR4, CCL20/CCR6, and CCL21/CCR7 pairs has been shown by Western blot analysis.8,15,19 To further accumulate clinical data and validate the potential attribution of these chemokine/chemokine receptor pairs in the prognosis of primary and metastatic liver tumors, we comparatively investigated expression profiles of CXCL12/CXCR4, CCL20/CCR6, and CCL21/CCR7 in HCC and CRLM specimens and their corresponding adjacent non-cancerous liver tissues, and elucidated their impact on the carcinogenesis and prognosis.
Methods
Patients and Samples
The present study was approved by the clinical research ethics committee of Tianjin Third Central Hospital, and written informed consent was obtained from all patients undergoing surgical resection in our hospital between 2007 and 2011. Surgical specimens and the corresponding non-cancerous tissue from the same samples were collected from patients with CRLM (n=11) and HCC (n=29) who underwent surgical resection in the Department of Hepatobiliary Surgery of Tianjin Third Central Hospital in China between 2007 and 2011. The clinical data and patient characteristics of all the patients were obtained prospectively from the clinical and pathological records and are summarized in Table 1.
Table 1.
Clinical Characteristics of Patients With HCC and CRLM.
| Variable | HCC (n=29) | CRLM (n=11) | p |
|---|---|---|---|
| Gender (male/female) | 24/5 | 5/6 | 0.018 |
| Age (mean ± SD) | 55.9 ± 7.57 | 55.73 ± 10.60 | 0.834 |
| Tumor size (≤3/>3 cm) | 6/23 | 5/6 | 0.117 |
| Tumor number (1/≥2) | 21/8 | 5/6 | 0.110 |
| Tumor grade (WD/MD/PD) | 8/8/13 | 3/4/4 | 0.844 |
| Follow-up time (months) | 63.50 (59.50, 76.50) | 24 (7, 92) | 0.350 |
Abbreviations: HCC, hepatocellular carcinoma; CRLM, colorectal liver metastases; WD, well differentiated; MD, moderately differentiated; PD, poorly differentiated.
Patient Follow-up
The follow-up period was defined as the time from the date of surgery to the date of patient death or the last follow-up point. All patients were monitored after surgery.
Immunohistochemistry
Formalin-fixed, paraffin-embedded tissues were cut into 4-μm-thick slices. Briefly, the slices were deparaffinized and rehydrated, and then autoclaved at 121C for 5 min in citrate buffer (10 mM sodium citrate, pH 6.0) for antigen retrieval. Hydrogen peroxide (3%) was used to block endogenous peroxidase activity of the tissue sections. The full list of the primary antibodies used is given in Table 2. Goat anti-mouse secondary antibodies (Zhongshan Golden Bridge Bio-tech, Beijing, China) were used for the three chemokines and their receptor staining. Immunohistochemical staining was performed according to the manufacturer’s instructions. 3, 3′-Diaminobenzidine tetrachloride was used as the chromogen, and cell nuclei were counterstained with hematoxylin and then sections were dehydrated, cleared, and mounted. For negative controls, the primary antibodies were replaced with normal rabbit immunoglobulin (IgG): sc 2027 (Santa Cruz Biotechnology, Inc.).
Table 2.
Antibody Information.
| Antigen | Antibody | Clonality | Species | Catalog No | Dilution | Source | Antigen Retrieval Condition |
|---|---|---|---|---|---|---|---|
| CXCL12 | Anti-SDF1 antibody | Polyclonal | Rabbit | ab9797 | 1:100 | Abcam, Inc. | 121C for 5 min in citrate buffer (10 mM sodium citrate, pH 6.0) |
| CXCR4 | Anti-human/mouse CD184 (CXCR4) | Polyclonal | Rabbit | 14-6009 | 1:100 | eBioscience, Inc. | the same as above |
| CCL20 | Anti-MIP3α antibody | Polyclonal | Rabbit | ab9829 | 1:50 | Abcam (Hong Kong) Ltd. | the same as above |
| CCR6 | Anti-CCR6 antibody | Polyclonal | Rabbit | ab109703 | 1:500 | Abcam (Hong Kong) Ltd. | the same as above |
| CCL21 | Anti-CCL21 antibody | Polyclonal | Rabbit | ab9851 | 1:1000 | Abcam, Inc. | the same as above |
| CCR7 | Anti-human CCR7 | Monoclonal | Rat | 14-1979 Clone3D12 |
1:100 | eBioscience, Inc. | the same as above |
Evaluation of Immunohistochemical Staining
The immunohistochemical staining of the three chemokine/chemokine receptor pairs in cells was observed with an optical microscope (BX51, Olympus, Tokyo, Japan) by two experienced independent observers who were blinded to the clinical data. Staining was graded according to the intensity and percentage of positive tumor cells as previously published.10 Briefly, high expression was defined as more than 30% of tumor cells showing strong or moderate staining. To avoid possible false positivity, the necrotic areas and edges of tissue sections were not included in the counting of immunohistochemical analyses.
Statistical Analysis
All statistical analyses were performed using SPSS 16.0 software. Quantitative data were expressed as median (25th and 75th percentiles) or mean ± SD. Differences between CRLM and HCC groups or tumor tissues and adjacent non-cancerous tissues were analyzed by the chi-square test and Fisher’s exact test. The Spearman correlation test was used to analyze the correlation between chemokine/chemokine receptor expression and clinicopathological parameters. Kaplan–Meier methods with the log-rank test were used to estimate differences in survival curves. Univariate and multivariate Cox proportional hazard models were used to determine the importance of covariates. Estimates for hazard ratios were reported with 95% confidence intervals. The p values<0.05 were considered to be statistically significant.
Results
Chemokine/Chemokine Receptor Expression in HCC and CRLM Tissues
Immunohistochemical staining of CXCL12 and CXCR4 proteins mainly showed staining in the cytoplasm of cancer and normal stromal liver cells of HCC and CRLM specimens (Fig. 1). Staining of CXCR4 was also observed sporadically in the mesenchymal elements of adjacent non-cancerous tissues of HCC specimens (Fig. 1E) and in endothelial cells and spindle cells of HCC tissues (Fig. 1J). Immunohistochemical staining of CCL20 and CCR6 proteins is shown in Fig. 2A to F. Positive signals of the CCL20 protein were observed mainly in the cytoplasm of cancer cells and normal stromal liver cells of HCC tissues (Fig. 2B). The CCR6 protein was observed mainly in the cell membrane of cancer cells and normal stromal liver cells of HCC tissues (Fig. 2D). In the CRLM specimens, the membrane of hepatocytes along the tumor invasion front (Fig. 2E) and the nuclear compartment of the CRLM cells (Fig. 2F) revealed immunoreactivity for CCR6. Cytoplasm and/or nuclear staining was identified for CCL21, while cytoplasm staining alone was observed for CCR7 in HCC and CRLM specimens (Fig. 3).
Figure 1.
CXCL12 and CXCR4 protein expression in representative HCC, CRLM, and adjacent non-cancerous tissues. Weak to intermediate immunostaining of the CXCL12 protein in the adjacent non-cancerous samples (A) and tumor samples of HCC (B). Intermediate immunostaining of CXCL12 in the adjacent non-cancerous samples (C) and tumor samples of CRLM (D). (E) Weak CXCR4 reactivity in the adjacent non-cancerous tissues of HCC. (F) Medium to intense CXCR4 reactivity in HCC specimens. (G) Weak to intermediate CXCR4 reactivity in the adjacent non-cancerous tissues of CRLM specimens. (H) Medium to intense CXCR4 reactivity in CRLM specimens. (I) The negative control of the adjacent non-cancerous tissues of HCC. (J) Intense CXCR4 reactivity in endothelial cells and spindle cells of HCC tissues (scale bars = 50 μm). Abbreviations: HCC, hepatocellular carcinoma; CRLM, colorectal liver metastases; NT, adjacent non-cancerous tissues; T, tumor tissues.
Figure 2.
CCL20 and CCR6 protein expression in representative HCC and CRLM tissues. (A) Most non-tumor samples showed no substantial CCL20 reactivity. (B) Weak to intermediate immunostaining of CCL20 in HCC specimens. (C) Weak to no substantial CCR6 reactivity in the adjacent non-cancerous tissues of HCC. (D) Medium to intense CCR6 reactivity in HCC. (E) Intense CCR6 reactivity in a streak of hepatocytes along the tumor invasion front in CRLM specimens. (F) Intense CCR6 reactivity in CRLM specimens. The negative control of non-tumor tissues (G) and HCC tissues (H) (scale bars = 50 μm). Abbreviations: HCC, hepatocellular carcinoma; CRLM, colorectal liver metastases; NT, adjacent non-cancerous tissues; T, tumor tissues.
Figure 3.
CCL21 and CCR7 protein expression in representative HCC and CRLM tissues. Weak immunostaining of CCL21 in HCC specimens (A) and high staining intensities of CCL21 in CRLM specimens (B). Weak to medium intensities of cytoplasmic signals of CCR7 in HCC specimens (C) and medium to high staining intensities of CCR7 in CRLM specimens (D). The negative control of HCC tissue (E) and CRLM tissue (F) (scale bars = 50 μm). Abbreviations: HCC, hepatocellular carcinoma; CRLM, colorectal liver metastases.
As shown in Table 3, the positive expression rate of CCL20 was 66% (19 of 29) in HCC tissues versus 31% (9 of 29) in matched adjacent non-cancerous tissues (p=0.017), while the positive expression rate of CCR6 was 72% (21 of 29) in HCC tissues versus 17% (five of 29) in matched adjacent non-cancerous tissues (p<0.001). No difference was observed in the expression levels of the chemokine/chemokine receptor pairs CXCL12/CXCR4 and CCL21/CCR7 between the HCC tissues and the adjacent non-cancerous tissues (p>0.05). Further analysis of the chemokine/chemokine receptor pairs expression level in the patients with CRLM showed no difference between the tumor tissues and the adjacent non-cancerous tissue (p>0.05). Furthermore, we analyzed the expression of these chemokine/chemokine receptor pairs in CRLM tissues versus HCC tissues. As shown in Table 4, the expression rate of CCL21 and CCR7 (CCL21: 100% [11 of 11] versus 55% [16 of 29]; CCR7: 100% [11 of 11] versus 66% [19 of 29]) was higher in patients with CRLM than in patients with HCC (p=0.007, 0.038). No difference was found in the expression levels of the chemokine/chemokine receptor pairs CXCL12/CXCR4/ and CCL20/CCR6 in CRLM and HCC tissues.
Table 3.
Differences in Chemokines/Chemokine Receptors Expression Levels in Tumor Tissues Versus Tumor Neighboring Tissues.
| HCC | CRLM | |||||
|---|---|---|---|---|---|---|
| NT | T | p | NT | T | p | |
| CXCL12 expression (−/+) | 12/17 | 11/18 | 1.000 | 3/8 | 6/5 | 0.193 |
| CXCR4 expression (−/+) | 12/17 | 7/22 | 0.263 | 5/5 | 4/7 | 0.500 |
| CCL20 expression (−/+) | 20/9 | 10/19 | 0.017 * | 5/6 | 5/6 | 1.000 |
| CCR6 expression (−/+) | 24/5 | 8/21 | 0.000 ** | 3/8 | 2/9 | 0.500 |
| CCL21 expression (−/+) | 19/10 | 13/16 | 0.186 | 2/9 | 0/11 | 0.238 |
| CCR7 expression (−/+) | 9/20 | 10/19 | 1.000 | 1/10 | 0/11 | 0.500 |
Abbreviations: HCC, hepatocellular carcinoma; CRLM, colorectal liver metastases; NT, adjacent non-cancerous tissues; T, tumor tissues.
p<0.05 was considered statistically significant (bold).
p<0.01 was considered significantly statistically significant (bold).
Table 4.
Differences in Chemokine/Chemokine Receptor Expression Levels in HCC Tissues Versus CRLM Tissues.
| HCC (n=29) | CRLM (n=11) | p | |
|---|---|---|---|
| CXCL12 expression (−/+) | 11/18 | 6/5 | 0.343 |
| CXCR4 expression (−/+) | 7/22 | 4/7 | 0.455 |
| CCL20 expression (−/+) | 10/19 | 5/6 | 0.522 |
| CCR6 expression (−/+) | 8/21 | 2/9 | 0.696 |
| CCL21 expression (−/+) | 13/16 | 0/11 | 0.007 ** |
| CCR7 expression (−/+) | 10/19 | 0/11 | 0.038 * |
Abbreviations: HCC, hepatocellular carcinoma; CRLM, colorectal liver metastases.
p<0.05 was considered statistically significant (bold).
p<0.01 was considered significantly statistically significant (bold).
Correlation Between Chemokine/Chemokine Receptor Expression Levels and Clinicopathological Factors of Patients With CRLM and HCC
The analysis of associations between chemokine/chemokine receptor expression levels and clinicopathological factors (e.g., gender, age, tumor size, tumor number, tumor grade, and 5-year survival rate) indicated that there was no statistically significant correlation in patients with CRLM (p>0.05, data not shown), while in the HCC group, CCR6 expression was negatively correlated with 5-year estimated disease-specific survival rates (r = −0.0373, p=0.046; Table 5).
Table 5.
Correlation Between Chemokine/Chemokine Receptor Expression and Clinicopathological Parameters of HCC Patients.
| Variable | CXCL12 | CXCR4 | CCL20 | CCR6 | CCL21 | CCR7 | ||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| r | p | r | p | r | p | r | p | r | p | r | p | |
| Gender (male/female) | 0.169 | 0.382 | 0.044 | 0.820 | −0.053 | 0.785 | −0.127 | 0.512 | −0.139 | 0.471 | −0.245 | 0.200 |
| Age (<55/≥55) | −0.276 | 0.147 | −0.022 | 0.908 | 0.075 | 0.690 | 0.064 | 0.741 | 0.024 | 0.901 | 0.075 | 0.697 |
| Tumor size (≤3/>3 cm) | −0.224 | 0.243 | 0.110 | 0.571 | 0.346 | 0.066 | −0.125 | 0.509 | 0.053 | 0.784 | −0.012 | 0.949 |
| Tumor number (1/≥2) | −0.154 | 0.427 | −0.012 | 0.949 | −0.039 | 0.840 | −0.137 | 0.479 | 0.246 | 0.198 | 0.123 | 0.525 |
| Tumor grade (WD/MD/PD) | 0.178 | 0.356 | −0.200 | 0.270 | −0.237 | 0.215 | −0.124 | 0.523 | −0.342 | 0.069 | 0.033 | 0.867 |
| 5-year survival rate | 0.109 | 0.574 | −0.058 | 0.757 | 0.269 | 0.158 | −0.373 | 0.046 * | 0.184 | 0.338 | 0.070 | 0.718 |
Abbreviations: HCC, hepatocellular carcinoma; WD, well differentiated; MD, moderately differentiated; PD, poorly differentiated.
p<0.05 was considered statistically significant (bold).
Predictors of Prognosis of Patients With HCC and CRLM After Resection
Univariate and multivariate analyses revealed that the expression levels of CXCR4 and CCR6 were independently associated with the overall survival of patients with HCC (Table 6). No factor was associated with the overall survival of patients with CRLM after hepatic resection (data not shown). Furthermore, Kaplan–Meier analysis was used to estimate the overall survival curves of negative and positive expression of the chemokine/chemokine receptor pairs in the resected hepatic tissues of patients with HCC and CRLM. In the HCC group, the expression of only CCR6 was associated with overall survival (the results of CCR6 are shown in Fig. 4, and the results of other chemokine/chemokine receptors are not given). In patients with CRLM, no factors were found to be associated with overall survival (data not shown).
Table 6.
Univariate and Multivariate Analyses of Clinicopathological Parameters Associated With Overall Survival in Patients With HCC.
| Factors | Univatiate | Multivariate | |
|---|---|---|---|
| p | HR (95% CI) | p | |
| Gender (male/female) | 0.832 | 0.355 (0.039–3.278) | 0.361 |
| Age (<55/≥55) | 0.816 | 0.881 (0.137–5.666) | 0.894 |
| Tumor size (≤3/>3 cm) | 0.947 | 1.909 (0.248–14.684) | 0.535 |
| Tumor number (1/≥2) | 0.216 | 0.634 (0.114–3.531) | 0.603 |
| Tumor grade (WD/MD/PD) | 0.382 | — | 0.559 |
| CXCL12 expression (−/+) | 0.183 | 0.347 (0.042–2.907) | 0.329 |
| CXCR4 expression (−/+) | 0.885 | 0.079 (0.008–0.773) | 0.029 * |
| CCL20 expression (−/+) | 0.273 | 1.197 (0.208–6.887) | 0.840 |
| CCR6 expression (−/+) | 0.016 * | 23.189 (2.351–228.736) | 0.007 ** |
| CCL21 expression (−/+) | 0.812 | 0.391 (0.075–2.033) | 0.264 |
| CCR7 expression (−/+) | 0.329 | 0.113 (0.011–1.125) | 0.063 |
Abbreviations: HCC, hepatocellular carcinoma; HR, hazard ratio; 95% CI, 95% confidence interval; WD, well differentiated; MD, moderately differentiated; PD, poorly differentiated.
p<0.05 was considered statistically significant (bold).
p<0.01 was considered significantly statistically significant (bold).
Figure 4.
Correlations between CCR6 expression and the overall survival rate in patients with HCC. Abbreviation: HCC, hepatocellular carcinoma.
Discussion
Previous studies have demonstrated that chemokines and their receptors present in the microenvironment of HCC influence the development of HCC through several aspects, including inflammation, effects on immune cells, angiogenesis, and direct effects on tumor cells.3–6 In the present study, we report the elevated expression of CCL20/CCR6 in HCC tissues compared with that in paired adjacent non-cancerous tissues. In addition, the expression levels of CCR6 were significantly correlated with the 5-year survival rate; this suggests that CCR6 expression may be of clinical relevance in the aggressiveness of HCC. Moreover, univariate and multivariate analyses demonstrated that CCR6 was an independent risk factor for predicting short overall survival in patients with HCC; this finding suggests that CCR6 could serve as a prognostic marker for HCC. These data agree well with a number of previous findings regarding the potential role of the CCL20/CCR6 system in the occurrence and progression of HCC. Previous studies have reported that the chemokine CCL20 and its receptor CCR6 play an important role in the tumor-chemokine network and promote tumor progression.23,24 Liu et al.12 revealed that the upregulation of CCR6 enhanced cell proliferation and carcinogenesis in HCC cells by regulating the expression of p21, p27, and cyclin-D1, and Li et al.25 reported that the CCL20/CCR6 cytokine network facilitates Treg activity in advanced grades and metastatic variants of HCC.
Recently, the CCL21/CCR7 axis has gained increasing attention because of its vital role in determining lymph node metastasis in various malignant processes and tumor types such as leukemia, melanoma, gastric, or non-small cell lung cancer, as well as its strong chemotactic effects on malignant processes. In our study, we found a significantly higher expression level of CCL21/CCR7 in patients with CRLM than in patients with HCC; this finding suggests an association between CCL21/CCR7 expression and the promotion of colorectal liver metastasis. Various studies have demonstrated that chemokines selectively expressed in certain tissues can promote metastasis by attracting specific CCR-expressing tumor cells and/or by providing growth stimulatory signals. Muller et al.21 demonstrated that the CCR7 chemokine receptor was involved in the metastasis of human breast cancer cells that express and secrete their respective CCL21 ligand to distant organs. On the basis of this concept, we speculate that the increased expression levels of CCR7 in CRC cells facilitate chemoattraction of CRC cells to the liver where the expression of CCL21 is high. To further verify this hypothesis and investigate the potential mechanisms, the analysis of the expression levels of the CCL21/CCR7 axis in the primary CRC sites and their roles in CRC cell proliferation and invasion is required.
The CXCL12/CXCR4 axis is considered as a critical factor that regulates tumor growth and progression during HCC and serves as a predictor of CRLM.23,26–28 Previous studies have revealed higher expression of CXCL12 and CXCR4 in HCC specimens than in the surrounding tissues.27 Different studies have demonstrated the important role of CXCR4 and CXCL12 in HCC metastasis and invasiveness of the tumor.23,28 A significant correlation was observed between CXCR4 expression and tumor progression, metastasis, and a decreased survival rate.17 However, the lack of a loss-of-function mutation of the tumor suppressor gene p53 on CXCR4 expression in HCC indicated yet another unidentified mechanism.29 We observed no correlation between CXCL12/CXCR4 expression and the development of HCC or CRLM. Similarly, CXCL12/CXCR4 expression showed no significant difference between tumor tissues and adjacent non-cancerous tissues in either cancer type. It may well be possible that the small sample size has led to this statistical bias; therefore, this result needs to be validated by a study of a larger sample size. In addition, the results of the survival analysis demonstrated significant associations of the chemokine/chemokine receptor pairs with survival, suggesting CXCR4 as a prognostic factor for positive outcome of patients with HCC. Various studies have suggested that the secretion of CXCL12/CXCR4 in tumors may mediate T-cell-dependent antitumor responses, which in turn exert antitumor effect.30–32
The identification of key targets promoting disease progression and metastasis is of great interest for the development of specific treatment strategies. Inhibition of the progression and metastasis of tumor cells by interfering with chemokine/chemokine receptor interactions is a promising new treatment strategy. Thus, targeting the CCL20/CCR6 axis may represent a potential treatment strategy and a new alternative for prognosis in HCC; moreover, the CCL21/CCR7 axis may serve as a predictive factor for the occurrence of liver metastases.
Supplemental Material
Supplemental material, DS_10.1369_0022155418824274 for The Diagnostic Value of Chemokine/Chemokine Receptor Pairs in Hepatocellular Carcinoma and Colorectal Liver Metastasis by Xiaolei Jiao, Guiming Shu, Hui Liu, Qin Zhang, Zhe Ma, Chaoyi Ren, Hongsheng Guo, Jingxiang Shi, Junguo Liu, Chuanshan Zhang, Yijun Wang and Yingtang Gao in Journal of Histochemistry & Cytochemistry
Acknowledgments
The authors thank the nursing staff of the Biobank of Tianjin Third Central Hospital for their cooperation in the collection of tissue samples and data.
Footnotes
Author Contributions: XJ and GS contributed to the experiment design, data analyses, and manuscript writing. HL, QZ, ZM, and CZ were responsible for the immunohistochemistry studies and image acquisition. CR, HG, JS, JL, and YW treated patients and collected material and clinical data. YG conceived of the study, data analysis and reviewed the manuscript. All authors read and approved the final manuscript.
Competing Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.
Funding: The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: This study was supported by grants from the Key Research Project on Basic Science of Tianjin Health Bureau (No. 2011KR07) and the Key Research Project of Tianjin Health Bureau (Nos. 15KG113 and 16KG150).
Contributor Information
Xiaolei Jiao, Artificial Cell Engineering Technology Research Center of Public Health Ministry, Tianjin Key Laboratory of Artificial Cells, Tianjin Institute for Hepatobiliary Disease, Tianjin Third Central Hospital, Tianjin, China.
Guiming Shu, Department of Hepatobiliary Surgery, Tianjin Third Central Hospital, Tianjin, China.
Hui Liu, Artificial Cell Engineering Technology Research Center of Public Health Ministry, Tianjin Key Laboratory of Artificial Cells, Tianjin Institute for Hepatobiliary Disease, Tianjin Third Central Hospital, Tianjin, China.
Qin Zhang, Department of Pathology, Tianjin Third Central Hospital, Tianjin, China.
Zhe Ma, Department of Pathology, Tianjin Third Central Hospital, Tianjin, China.
Chaoyi Ren, Department of Hepatobiliary Surgery, Tianjin Third Central Hospital, Tianjin, China.
Hongsheng Guo, Department of Hepatobiliary Surgery, Tianjin Third Central Hospital, Tianjin, China.
Jingxiang Shi, Department of Hepatobiliary Surgery, Tianjin Third Central Hospital, Tianjin, China.
Junguo Liu, Department of Hepatobiliary Surgery, Tianjin Third Central Hospital, Tianjin, China.
Chuanshan Zhang, Department of Pathology, Tianjin Third Central Hospital, Tianjin, China.
Yijun Wang, Department of Hepatobiliary Surgery, Tianjin Third Central Hospital, Tianjin, China.
Yingtang Gao, Artificial Cell Engineering Technology Research Center of Public Health Ministry, Tianjin Key Laboratory of Artificial Cells, Tianjin Institute for Hepatobiliary Disease, Tianjin Third Central Hospital, Tianjin, China.
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Supplementary Materials
Supplemental material, DS_10.1369_0022155418824274 for The Diagnostic Value of Chemokine/Chemokine Receptor Pairs in Hepatocellular Carcinoma and Colorectal Liver Metastasis by Xiaolei Jiao, Guiming Shu, Hui Liu, Qin Zhang, Zhe Ma, Chaoyi Ren, Hongsheng Guo, Jingxiang Shi, Junguo Liu, Chuanshan Zhang, Yijun Wang and Yingtang Gao in Journal of Histochemistry & Cytochemistry