沉默c2orf68基因对结直肠癌细胞增殖的影响

Effect of Silencing c2orf68 Gene on the Proliferation of Colorectal Adenocarcinoma Cells

目的:通过RNA干扰技术抑制人结直肠癌SW480、SW620及LS174T细胞株中c2orf68基因的表达,进一步观察其生物学特性的改变,以阐明c2orf68基因对结直肠癌细胞增殖的影响。方法:利用siRNA在线设计软件设计合成干扰片段,并将2个干扰片段转入所试细胞株中。应用RT-PCR法检测细胞mRNA的表达;利用MTT实验观察细胞增殖情况;应用流式细胞技术检测细胞凋亡。结果:转染siRNA片段后,SW480、SW620及LS174T细胞株的mRNA表达都下降,其抑制率分别为:50.05%、44.79%;49.69%、30.00%;50.15%、45.79%。随后,选取干扰效率高的siRNA1干扰片段做后续实验。MTT实验发现,siRNA转染LS174T细胞后,细胞的增殖能力明显下降,细胞的增殖抑制率为21.2%(P<0.05)。同时,采用流式细胞仪检测细胞的凋亡,实验表明,实验组细胞凋亡率为18.13%(P<0.05)。结论:成功筛选了抑制结直肠癌c2orf68基因的特异性干扰片段,该片段可以下调人结直肠癌细胞株中c2orf68的mRNA,抑制人结直肠癌细胞的生长,促进细胞凋亡,说明该基因可能与结直肠癌的细胞增殖有关,进一步调控相应生物学特性的改变,引起结直肠癌的发生。

Objective ：To explore the c2o0C68 gene expression of human colorectal adenocarcinoma SW480, SW620 and LS174T cell lines. Further more, to observe the change of their biological characteristics in three cell lines by applying RNA interference technique to silence c2o（9C68 and to clarify its in the pathogenesis of colorectal cancer. Methods：Using siRNA online design software designed and synthesised of siRNA interference fragments, and transported them to the three cell lines. The mRNA expression of the c2o0C68 were detected by RT-PCR, the cell proliferation was checked by MqT, and the cell apoptosis was measured by flow-cytometry. Results： The mRNA expression of the c2orf68 was inhibited by the two pairs siRNA in SW480, SW620 and LS174T cell lines. It showed that the inhibition rates were as follows ： 50.05% ,50.05% ; 49.69% ,49.69% ; 50.15% ,45.79%. Then, siRNA1 of high interference efficiency was selected. The cell proliferation inhibitory rate was 21.2% in LS174T cell line（P 〈0.05）. The cell apoptosis rate was 18.13% in LS174T cell line（P 〈0.05）. Conclusions： The successfully obtained siRNA fragement which specifically degraded its c2orf68 mRNA, inhibited the growth of the test cells, promoted cell apoptosis. It indicated that c2orf68 might be associated with the proliferation on colorectal cancer cells, its pathogenesis might be through gene expression, further regulated the change of the corresponding biological characteristics, and caused the occurrence of colorectal cancer.