干扰CHD1L基因对肝内胆管细胞癌细胞系增殖和侵袭的影响

李师淼; 花京文; 袁汀浩; 丁炎宝; 熊虎; 李洁; 曹青; 黄长文

doi:10.3971/j.issn.1000-8578.2017.17.0014

干扰CHD1L基因对肝内胆管细胞癌细胞系增殖和侵袭的影响

CHD1L Silencing Inhibits Proliferation and Invasion of Intrahepatic Cholangiocarci-noma Cell Lines

摘要

摘要:
目的探讨沉默CHD1L基因对肝内胆管细胞癌（ICC）细胞系增殖和侵袭的影响。
方法采用Western blot和qRT-PCR法检测三种ICC细胞系（RBE、HCCC-9810、HUCCT1）中CHD1L蛋白和mRNA的表达；选择CHD1L高表达的ICC细胞系，将CHD1L-RNAi转染该细胞后，用Western blot法筛选出干扰效果最好的CHD1L-RNAi；应用CCK-8法、平板克隆集落形成实验、细胞划痕实验和Transwell迁移和侵袭实验检测转染CHD1L-RNAi后细胞增殖、迁移和侵袭能力的变化。
结果 CHD1L蛋白和mRNA水平在RBE细胞系中明显高于HCCC-9810和HUCCT1细胞系（P=0.003, P=0.012和P=0.008, P=0.015）；CHD1L-RNAi1的干扰效果明显优于CHD1L-RNAi2和CHD1L-RNAi3（P=0.003, P=0.008）；干扰CHD1L的表达，结果显示CHD1L-RNAi1组的增殖率、集落形成数、迁移率和穿膜的细胞数明显低于未处理组和阴性对照组，差异有统计学意义（P < 0.05）。
结论干扰CHD1L基因能有效抑制RBE细胞系的增殖、迁移及侵袭能力。

Abstract:
Objective To investigate the effect of CHD1L silencing by RNAi on the growth and invasion of intrahepatic cholangiocarcinoma(ICC) cells line.
Methods The protein and mRNA expressions of CHD1L in three types of ICC cell lines, RBE, HCCC-9810 and HUCCT1, were determined by Western blot and qRT-PCR. The overexpression of CHD1L cell line was selected and after transfection with CHD1L-RNAi, the protein expressions of CHD1L were measured by Western blot. CCK-8 assay and plate clone formation assay were used to define the cell proliferation rate. Cell migration and invasion was detected by cell scratch assay and Transwell migration and invasion assay.
Results The protein and mRNA expressions of CHD1L in RBE cell line were significantly higher than those in HCCC-9810, HUCCT1 cell lines (P=0.003, P=0.012 and P=0.008, P=0.015). After CHD1L-RNAi was transfected into RBE cell line, we found that the interference effect of CHD1L-RNAi1 was significantly better than those of CHD1L-RNAi2 and CHD1L-RNAi3 (P=0.003, P=0.008). CCK-8 results showed CHD1L-RNAi1 group could obviously inhibit the proliferation of RBE cells line (P < 0.01). Meantime, the results of plate clone formation assay, cell scratch and Transwell migration and invasion experiment showed the number of colony formation, the rate of migration, the cells number of migration and invasion of CHD1L-RNAi1 group were significantly lower than those in normal and negative control groups (P < 0.05).
Conclusion C H D1L silencing by CHD1L-RNAi could inhibit the proliferation, migration and invasion of RBE cells line.

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