RhoC—siRNA表达载体的构建及其对肝癌细胞体外侵袭能力的影响

Construction of RhoC-siRNA expression vector and its role in invasion and metastasis ability of hepatoma carcinoma cell

目的构建RhoC-siRNA表达载体，研究其对肝癌细胞体外侵袭潜能的影响。方法以脂质体转染法稳定转染人SK-Hepl肝癌细胞株，Western印迹鉴定转染细胞中RhoC蛋白表达的抑制情况，观察转染前后细胞黏附、运动及体外侵袭能力的改变。结果酶切及测序证实RhoC—siRNA真核表达载体构建成功。通过Western印迹检测证实，转染RhoC—siRNA表达载体的肝癌细胞中RhoC蛋白表达抑制达60％，其黏附、移动及体外侵袭能力均降低。结论RhoC—siRNA表达载体能有效抑制RhoC在肝癌细胞株SK—Hepl中的表达，同时抑制细胞的侵袭转移潜能，为肝癌的生物学治疗提供新的方法。

Objective To construct a RhoC-siRNA expression vector and study its role in invasion and metastasis ability of hepatoma carcinoma cells （HCC）. Methods RhoC-siRNA gene was synthesized and cloned into the expression vector pSilencer 2. 1. The constructed RhoC-siRNA expression Vector was stably transfected into SK-Hepl of HCC. By using Western blotting, the inhibitive effect of RhoC-siRNA on expression of RhoC in transfected HCC was detected, the ability of cell adhesion, cell migration, and cell invasion before and after transfection were observed too. Results Enzyme digestion and DNA sequencing confirmed that the RhoC specific siRNA expression vector was constructed successfully. After transfection, the RhoC expression was inhibited by 60% , and the ability of cell adhesion, cell migration, and cell invasion was decreased markedly, which was proved by Western blotting. Conclusions RhoC-siRNA expression vector can effectively suppress RhoC expression in SK-Hepl of HCC and decrease potential of cell invasion and cell metastasis, which may provide a novel applicable strategy for gene therapy of HCC.