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Effects of Livin gene silencing on biological characteristics of colon cancer LoVo cells
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Abstract:
Objective :To observe the effects of Livin on biology characteristics, such as proliferation and apoptosis, of colon cancer LoVo cells by RNA interference (RNAi) targeting Livin gene. Methods: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 targeting Livin gene were constructed and transfected into LoVo cells. Then the expression of Livin was detected by RT-PCR and Western blotting. And the apoptosis, cell cycle, colony formation, proliferation of LoVo cells, as well as sensitivity of LoVo cells to cisplatin, were detected by flow cytometry, colony formation assay, and MTT. Results: Interference vectors pSilencer4.1-L1 and pSilencer4.1-L2 were successfully constructed. pSilencer4.1-L1, but not pSilencer4.1-L2, effectively silenced the Livin mRNA and protein expression in LoVo cells (P<0.01). Compared with the control group, LoVo cells in pSilencer4.1-L1 transfection group showed an increased apoptosis rate (\[24.2±32\]% vs \[8.1±1.4\]%, P＜0.01), a decreased cell proliferation (inhibitory rate: about 70% after 72 h), a decreased colony formation rate (\[15±4.6\]% vs \[85±5.8\]%, P＜0.01), increased S phase cells (\[45.7±49\]% vs \[28.0±3.0\]%, P＜0.01), decreased G1 phase cells (\[43.0±5.2\]% vs \[62.8±5.1\]%, P＜0.01), and an increased sensitivity to cisplatin (apoptosis rate increased from \[43.4±6.9\]% to \[65.3±6.2\]%, P＜0.01). Conclusion: pSilencer4.1-L1 can effectively silence Livin gene expression in colon cancer LoVo cells, inhibit proliferation and colony formation, induce apoptosis, and enhance sensitivity of LoVo cells to cisplatin.
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Project Supported:
Project supported by the Science and Technology Development Foundation of Xi’an \[No. HM1124(2)\]
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