CXC型趋化因子配体10与12在胆囊癌组织中的表达水平及在肿瘤侵袭中的作用机制

Expression of CXCL10 and CXCL12 in gallbladder carcinoma and their mechanism of action in tumor invasion

目的探讨CXC型趋化因子配体10(CXCL10)和12在胆囊癌组织中的表达水平及其参与肿瘤侵袭的机制。方法选择2020年4月—2023年4月在中国人民解放军中部战区总医院手术切除的56例胆囊癌患者肿瘤组织及癌旁组织标本,使用RT-PCR法检测癌组织及癌旁组织CXCL10 m RNA、CXCL12 m RNA表达,分析患者癌组织中CXCL10 m RNA、CXCL12m RNA表达与患者临床病理参数的关系。使用人胆囊癌细胞株GBC-SD,构建CXCL10、CXCL12低表达胆囊癌细胞,CCK-8法检测CXCL10、CXCL12低表达对胆囊癌细胞株增殖的影响,Transwell检测CXCL10、CXCL12低表达对胆囊癌细胞株侵袭能力的影响,并采用Western Blot法检测胆囊癌细胞PI3K/Akt通路表达情况。计量资料两组间比较采用配对t检验或成组t检验;多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果在胆囊癌患者中,癌组织CXCL10、CXCL12 m RNA相对表达量均显著高于癌旁组织(1.857±0.315 vs 1.024±0.203、2.038±0.374 vs 1.064±0.221,t值分别为16.342、16.778,P值均<0.05)。不同TNM分期、有无淋巴结转移、有无远处转移以及不同肿瘤直径患者的CXCL10、CXCL12 m RNA比较差异均有统计学意义(P值均<0.05)。CXCL10:si-CXCL10组细胞CXCL10 m RNA相对表达量、CXCL10蛋白表达、CCK-8吸光值、细胞迁移数量、p-PI3K及p-Akt蛋白表达均显著低于对照组和si-RNA组(P值均<0.05);CXCL12:si-CXCL12组细胞CXCL12 m RNA相对表达量、CXCL12蛋白表达、CCK-8吸光值、细胞迁移数量、p-PI3K及p-Akt蛋白表达均显著低于对照组和si-RNA组(P值均<0.05)。结论胆囊癌组织中的CXCL10及CXCL12表达升高,抑制CXCL10、CXCL12表达后,胆囊癌细胞增殖、侵袭受到明显抑制,其机制可能与抑制PI3K/Akt通路磷酸化有关。

Objective To investigate the expression levels of CXCL10 and CXCL12 in gallbladder carcinoma and their mechanism of action in tumor invasion.Methods Tumor tissue samples and adjacent tissue samples were collected from 56 patients with gallbladder carcinoma who underwent surgical resection in General Hospital of Central Theater Command from April 2020 to April 2023.RT-PCR was used to measure the mRNA expression levels of CXCL10 and CXCL12 in cancerous tissue and adjacent tissue,and the correlation of the mRNA expression levels of CXCL10 and CXCL12 in cancerous tissue with clinicopathological parameters was analyzed.The human gallbladder carcinoma cell line GBC-SD was used to construct gallbladder carcinoma cells with low expression of CXCL10 and CXCL12.CCK8 assay was used to observe the effect of low expression of CXCL10 and CXCL12 on the proliferation of gallbladder carcinoma cells,Transwell assay was used to observe the effect of low expression of CXCL10 and CXCL12 on the invasion ability of gallbladder carcinoma cells,and Western blot was used to measure the expression of the PI3K/Akt pathway in gallbladder carcinoma cells.The paired t-test or independent-samples t-test was used for comparison of measurement data between two groups;an analysis of variance was used for comparison between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results In the patients with gallbladder carcinoma,the relative mRNA expression levels of CXCL10 and CXCL12 in cancerous tissue were significantly higher than those in adjacent tissue(CXCL10:1.857±0.315 vs 1.024±0.203,t=16.342,P<0.05;CXCL12:2.038±0.374 vs 1.064±0.221,t=16.778,P<0.05).There were significant differences in the relative mRNA expression levels of CXCL10 and CXCL12 between the patients with different TNM stages,presence or absence of lymph node metastasis or distant metastasis,and tumor diameters(all P<0.05).Compared with the control group and the si-RNA group,the si-CXCL10 group had significantly lower relative mRNA and protein expression levels of CXCL10 and CXCL12,CCK-8 absorbance values,number of cell migration,and protein expression levels of p-PI3K and p-Akt(all P<0.05).Conclusion There are increases in the expression of CXCL10 and CXCL12 in gallbladder carcinoma tissue,and the proliferation and invasion of gallbladder carcinoma cells are significantly inhibited after inhibition of the expression of CXCL10 and CXCL12,which might be associated with the inhibition of the phosphorylation of the PI3K/Akt pathway.