Twist蛋白在结直肠癌中的表达水平及其促进癌症发展的机制研究

Expression of Twist protein in colorectal cancer and mechanism of its cancer promoting effect

摘要:

目的  研究Twist蛋白在结直肠癌中的表达水平，其对结直肠癌细胞的增殖以及侵袭能力的影响，并探讨其促进癌症发展具体的机制。方法  经结肠镜活检取20例患者组织和20例正常人的标本，采用实时定量PCR的方法检测了Twist的mRNA表达水平。采用脂质转染法对结直肠癌细胞系CT-26细胞进行RNA干扰，采用CCK-8法及transwell法观察干扰Twist后对结直肠癌细胞的增殖以及侵袭能力的影响；采用RT-PCR以及Western blot检测了CT-26细胞中MMP-9的mRNA及蛋白水平。结果  Twist在结直肠癌组织中的mRNA水平显著高于对应癌旁组织以及对照组（P <0.05）；在伴有淋巴结转移的癌组织中Twist mRNA的表达水平显著高于无淋巴结转移的组织（P <0.05）；与转染对照组比较，Twist siRNA转染的CT-26细胞增殖水平及迁徙能力均显著降低（P <0.05）；干扰Twist的CT-26细胞能够显著降低金属蛋白酶MMP-9 mRNA及蛋白的水平。结论  Twist蛋白能够增加结直肠癌癌细胞的增殖及迁徙能力，其影响机制可能与金属蛋白酶MMP-9有关。

Abstract:

Objective To explore the expression of Twist protein in colorectal cancer and its impact on the pro-liferation and invasion of colorectal cancer cells, and to clarify the mechanism of its cancer promoting effect. Methods Twenty tissue specimens were selected from colorectal cancer patients and normal people through colonoscopy biopsy, respectively, then the expression of Twist mRNA was detected through real time quantitative PCR method. The colorectal cancer cell line CT-26 cell was RNA interfered by lipid transfection method, and was observed the impact of Twist after interference on the proliferation and invasion of colorectal cancer cells by CCK-8 method and transwell method, and the expression of mRNA and protein of MMP-9 in CT-26 cell were detected by RT-PCR and Western blot method. Results The expression of Twist mRNA in colorectal cancer tissues was significantly higher than in para-carcinoma tissues the control group (P < 0.05). The expression of Twist mRNA in colorectal cancer tissues with lymph node metastasis was significantly higher than in colorectal cancer tissues without lymph node metastasis (P < 0.05). Compared with transfection control group, the proliferation and migration ability of CT-26 cells transfected by Twist siRNA were significantly decreased (P < 0.05). CT-26 cells interfering Twist reduced the expression of mRNA and protein of metal protease MMP-9. Conclusions Twist protein strengthens the proliferation and migration ability of colorectal cancer cells, and the mechanism may relate with metal protease MMP-9.