MUC1对人结肠癌细胞HCT116生物学行为的影响

Effect of MUC1 on Biological Behaviour of Human Colorectal Cancer Cell HCT116

目的:观察MUC1对人结肠癌细胞HCT116增殖、侵袭及化疗敏感性的影响。方法:采用MUC1表达阴性的结肠癌细胞株HCT116,通过慢病毒转染、嘌呤霉素筛选、半定量RT-PCR和Western blot鉴定构建稳定表达MUC1的HCT116细胞株;实验分空病毒组和MUC1病毒组;CCK实验和软琼脂克隆形成实验检测两组细胞的增殖能力,Transwell侵袭实验检测两组细胞的侵袭能力;MTT法和流式细胞仪检测两组细胞对奥沙利铂的敏感性,酶底物法检测Caspase-3活性。结果:获得稳定表达MUC1的HCT116细胞株;两组细胞贴壁生长无差异;与空病毒组相比,MUC1病毒组细胞软克隆形成数增加,穿过小室的细胞数增加(P<0.05);MUC1病毒组细胞对奥沙利铂的敏感性降低,MUC1病毒组Caspase-3活性水平低于空病毒组(P<0.05)。结论:MUC1与结肠癌的非锚定依赖生长、侵袭和化疗敏感性有关。

Objective： To investigate the effect of MUC1 on proliferation,invasion and chemosensitivity of human colorectal cancer cell line HCT116. Methods： The HCT116 human colon cancer cells were transfected with lentiviral vectors with and without MUC1,then selected by puromycin. The expression of MUC1 was detected by SqRT-PCR and Western blot. There were two groups in our experiment： MUC1 lentivirus group and control lentivirus group. CCK method and soft agar colony formation method were used to detect cells proliferation,Transwell membrane assay to detect cells invasion,MTT method and flow cytometry to detect cells sensitivity to oxaliplatin and the colorimetric method to detect Caspase-3 activity. Results： HCT116 cell line with stable expression of the MUC1 was acquired. Compared with control lentivirus group,the number of soft agar colonies in MUC1 lentivirus group was increased significantly（ P 〈 0. 05）,the number of invasive cells was increased significantly（ P 〈 0. 05）. Chemosensitivity was decreased in the MUC1 lentivirus group（ P 〈 0. 05）.Caspase-3 activity was significantly low in the MUC1 lentivirus group（ P 〈 0. 05）. Conclusion： MUC1 is related to the anchorage-independent growth,invasion and chemotherapy of colorectal cancer.