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首页> 外文期刊>World journal of gastroenterology : >Effect of p27mt gene on apoptosis of the colorectal cancer cell line Lovo.
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Effect of p27mt gene on apoptosis of the colorectal cancer cell line Lovo.

机译：p27mt基因对大肠癌细胞Lovo凋亡的影响。

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AIM: To construct p27mt recombinant adenovirus, transfect the colorectal cell line Lovo and observe the effects of p27mt on Lovo cell apoptosis and cell cycle inhibition. METHODS: We constructed recombinant adenovirus containing p27mt by homologous recombination in bacteria. The colorectal cancer cell line Lovo was infected with recombinant replication-defective adenovirus Ad-p27mt, and expression of p27mt was determined by Western blotting; the inhibitory effect of p27mt on Lovo cells was detected by cytometry. Cell cycle was determined by flow cytometry. DNA fragment analysis identified the occurrence of apoptosis. RESULTS: The recombinant adenovirus which already contained p27mt target gene was successfully constructed. When multiplicity of infection was >or= 50, the infection efficiency was 100%. After transfection of Lovo cells with Ad-p27mt the cells had high p27 expression which was identified by immunoblotting assay. PI staining and flow cytometry showed that 77.96% of colorectal cancer cells were inhibited in phase G(0)/G(1), while in the Ad-LacZ group and blank control group, 27.57% and 25.29% cells were inhibited in the same phase, respectively. DNA fragment analysis, flow cytometry and TUNEL assay demonstrated that p27mt is able to induce apoptosis in colorectal cancer cells. CONCLUSION: p27mt has an obvious blocking effect on colorectal cancer cell cycle, and most cells were inhibited in phase G(0)/G(1). Therefore, p27mt can induce apoptosis in colorectal cells.
机译：目的：构建p27mt重组腺病毒，转染大肠癌细胞Lovo，观察p27mt对Lovo细胞凋亡和细胞周期抑制的影响。方法：我们通过细菌中的同源重组构建了含有p27mt的重组腺病毒。用重组复制缺陷型腺病毒Ad-p27mt感染结直肠癌细胞系Lovo，并通过蛋白质印迹法测定p27mt的表达。细胞计数法检测p27mt对Lovo细胞的抑制作用。细胞周期通过流式细胞术确定。 DNA片段分析鉴定出凋亡的发生。结果：成功构建了已包含p27mt靶基因的重组腺病毒。当感染复数大于或等于50时，感染效率为100％。用Ad-p27mt转染Lovo细胞后，细胞具有高p27表达，可通过免疫印迹测定法进行鉴定。 PI染色和流式细胞仪显示，在G（0）/ G（1）期抑制了77.96％的结直肠癌细胞，而在Ad-LacZ组和空白对照组中，相同的时间抑制了27.57％和25.29％的癌细胞相分别。 DNA片段分析，流式细胞术和TUNEL分析表明p27mt能够诱导大肠癌细胞凋亡。结论：p27mt对结直肠癌细胞周期有明显的阻断作用，大多数细胞在G（0）/ G（1）期受到抑制。因此，p27mt可诱导大肠癌细胞凋亡。

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