钙黏蛋白12在结肠癌细胞株与结肠直肠癌组织中的表达及其影响

Expression of both cadherin-12 in colon cancer cell lines and colorectal cancer and the influence on carcinogenesis

目的:研究钙黏蛋白12(cadherin-12,CDH-12)在结肠癌细胞株与结肠直肠癌组织中的表达及其对结肠癌细胞株增殖、侵袭、迁移的影响。方法:应用实时PCR和Western印迹法在7株结肠癌细胞株中筛选CDH-12高表达细胞株;shRNA瞬转干扰高表达细胞株SW1116,Western印迹法验证干扰效果;CCK-8检测癌细胞增殖活性变化;transwell实验验证癌细胞迁移、侵袭能力的变化;组织芯片免疫组织化学技术检测CDH-12在结肠直肠癌组织中的表达。结果:实时PCR和Western印迹结果显示CDH-12在细胞株中表达,SW1116和LoVo两个细胞株高表达CDH-12,在成功下调SW1116细胞中CDH-12表达之后,SW1116结肠癌细胞株的增殖、迁移、侵袭能力较对照组明显下降;免疫组化染色结果显示,CDH-12在结肠直肠癌组织中的表达明显高于癌旁正常组织中的表达。结论:CDH-12在结肠直肠癌组织中的表达明显高于癌旁正常组织,证实了CDH-12为癌基因的可能性;下调CDH-12在结肠癌细胞株SW1116的表达可明显抑制细胞的增殖、迁移、侵袭能力,显示其在肿瘤细胞生长和转移中的重要作用。

Objective To investigate the expression of cadherin-12 in colon cancer cell lines and colorectal cancer and its influence on the carcinogenesis. Methods The cadhefin-12 high-expression cell line was chosen with screen of 7 cell lines by both real-time PCR and Western blot. ShRNA oligos targeting cadherin-12 was transfected into SWlll6 and the efficacy of eadherin-12 knockdown was verified by Western blot. The proliferation ability in SWlll6 after ShRNA transfection was determined by CCK-8 assay. The transwell assay was applied to test the migration and invasion ability. Cadherin-12 expression in colorectal cancer tissues were detected by immunohistochemistry. Results Real-time PCR and Western analysis were used to demonstrate that all the seven cell lines expressed cadherin-12. SWlll6 and LoVo were the top-two cell lines with high expression of cadherin-12. Compared to the control group, the ability of SWlll6 in proliferation, migration and invasion was obviously suppressed after shRNA transfection. The expression of cadherin-12 differed between the colorecta] cancer tissue and the adjacent normal tissue. Conclusions Expression of cadherin-12 was higher in colorectal cancer tissue. The knockdown of cadherin-12 expression in SW1116 cell line could obviously suppress the ability to proliferation, migration and invasion. Our results suggest an important role of cadherin-12 gene on the growth and metastasis in colorectal cancer.