反义RhoC基因抑制人QBC939胆管癌细胞的增殖和侵袭能力。

Antisense RhoC gene suppresses proliferation and invasion capacity of human QBC939 cholangiocarcinoma cells.

作者信息

Shi Zheng, Chen Ming-Liu, He Qing-Liang, Zeng Jin-Hua

机构信息

Department of General Surgery, First Affiliated Hospital, Fujian Medical University, Fuzhou 350005, China.

出版信息

Hepatobiliary Pancreat Dis Int. 2007 Oct;6(5):516-20.

PMID:17897917

Abstract

BACKGROUND

Cholangiocarcinoma is a highly aggressive, fatal malignancy, which is resistant to all current therapeutic approaches. The recent elevation in the incidence of cholangiocarcinoma has highlighted the need for novel approaches targeting the molecular basis of its invasiveness. Previously we reconstructed a RhoC antisense eukaryotic expression vector and transfected it into a cholangiocarcinoma cell line (QBC939) by the lipofectamine method. This study was undertaken to determine the effect of the antisense RhoC gene on the proliferation and invasion capacity of QBC939.

METHODS

Antisense RhoC cDNA was transfected into QBC939 with lipofectin 2000. The cell growth curve was constructed to determine the proliferation rate of cells; flow cytometry was used to analyze cell cycle changes of the tumor cells; and a Boyden chamber was used to assess the invasive ability of the cells before and after gene transfection.

RESULTS

After the antisense RhoC cDNA was transfected, the number of colonies formed was significantly lower than that in the other two groups (54+/-8 vs. 91+/-11 vs. 90+/-9, P<0. 05) so was the number of the cells which crossed to the lower surface of the matrigel-coater filters (36+/-6 vs. 96+/-12 vs. 95+/-7, P<0.05). There was also a higher percentage of transfected cells in G1 phase than in the other two groups (52.5% vs. 43.4% vs. 43.7%).

CONCLUSION

The antisense RhoC gene can suppress the capacities of proliferation and invasion in a cholangiocarcinoma cell line in vitro.

摘要

背景

胆管癌是一种侵袭性很强的致命恶性肿瘤，对目前所有的治疗方法均具有抗性。近期胆管癌发病率的上升凸显了针对其侵袭性分子基础的新方法的必要性。此前我们构建了RhoC反义真核表达载体，并通过脂质体法将其转染至胆管癌细胞系（QBC939）中。本研究旨在确定反义RhoC基因对QBC939增殖和侵袭能力的影响。

方法

用脂质体2000将反义RhoC cDNA转染至QBC939。绘制细胞生长曲线以确定细胞增殖率；采用流式细胞术分析肿瘤细胞的细胞周期变化；使用Boyden小室评估基因转染前后细胞的侵袭能力。

结果

转染反义RhoC cDNA后，形成的集落数量显著低于其他两组（54±8 vs. 91±11 vs. 90±9，P<0.05），穿过基质胶包被滤膜下表面的细胞数量也显著低于其他两组（36±6 vs. 96±12 vs. 95±7，P<0.05）。转染细胞处于G1期的百分比也高于其他两组（52.5% vs. 43.4% vs. 43.7%）。