miR-183在子宫内膜癌Ishikawa细胞中的表达及其对p53蛋白的调节作用

发布时间：2018-01-15 09:43

本文关键词：miR-183在子宫内膜癌Ishikawa细胞中的表达及其对p53蛋白的调节作用　出处：《中国妇幼保健》2016年24期 　论文类型：期刊论文

更多相关文章： 子宫内膜癌 Ishikawa细胞 miR- p蛋白

【摘要】：目的探讨miR-183在子宫内膜癌Ishikawa细胞中的表达及其对p53蛋白表达和细胞增殖、凋亡的影响。方法采用LipofectaminerTM2000脂质体将miR-183的特异性抑制剂反义寡核苷酸转染到Ishikawa细胞(实验组),不相关序列、脂质体和未转染组细胞分别为阴性对照组、脂质体对照组和空白对照组。采用CCK-8法和流式细胞仪分别检测各组子宫内膜癌Ishikawa细胞的增殖和凋亡情况;采用RT-PCR检测miR-183和p53 mRNA在各组子宫内膜癌Ishikawa细胞中的表达水平;采用Western blot检测p53蛋白在各组子宫内膜癌Ishikawa细胞中的表达水平。结果 CCK-8检测结果显示:转染0~24 h,实验组、空白对照组、脂质体对照组子宫内膜癌Ishikawa细胞增速相似,阴性对照组子宫内膜癌Ishikawa细胞增速略快;转染24~48 h,实验组、空白对照组和脂质体对照组子宫内膜癌Ishikawa细胞增速较快;转染48~72 h,实验组子宫内膜癌Ishikawa细胞较其余3组增长速度快;转染72~96 h,4组子宫内膜癌Ishikawa细胞继续保持增长状态,且达到对数生长期,实验组子宫内膜癌Ishikawa细胞较其余3组增速快。流式细胞学检测结果显示:转染3 d后,实验组较其余3组早期凋亡率明显降低,差异有统计学意义(P0.05);阴性对照组较其余3组早期凋亡率明显增高,差异有统计学意义(P0.05)。RT-PCR结果显示:实验组miR-183表达水平明显低于其余3组,差异有统计学意义(P0.05);实验组p53 mRNA表达水平明显低于其余3组,差异有统计学意义(P0.05)。Western blot检测结果显示:实验组p53蛋白表达水平明显高于其余3组,差异有统计学意义(P0.05)。结论 miR-183在子宫内膜癌Ishikawa细胞中有表达,miR-183抑制剂能有效抑制miR-183在子宫内膜癌Ishikawa细胞中的表达,促进细胞增殖,抑制细胞凋亡,上调p53蛋白表达。miR-183可能通过抑制p53蛋白表达而抑制子宫内膜癌细胞增殖,促进凋亡,从而影响子宫内膜癌的发生、发展及预后。
[Abstract]:Objective to investigate the expression of miR-183 in endometrial carcinoma Ishikawa cells and its expression of p53 protein and cell proliferation. Methods LipofectaminerTM2000 liposome was used to transfect antisense oligodeoxynucleotides (antisense oligonucleotides), a specific inhibitor of miR-183, into Ishikawa cells. Experimental group). Unrelated sequences, liposomes and untransfected cells were respectively negative control group. CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of endometrial carcinoma Ishikawa cells in each group. RT-PCR was used to detect the expression of miR-183 and p53 mRNA in Ishikawa cells of endometrial carcinoma. Western blot was used to detect the expression of p53 protein in Ishikawa cells of endometrial carcinoma. Results the results of CCK-8 assay showed that the expression of p53 protein was 0 24 h after transfection. In experimental group, blank control group and liposome control group, the growth rate of Ishikawa cells in endometrial carcinoma was similar, while that in negative control group was a little faster. After transfection for 24 ~ 48 h, the growth rate of Ishikawa cells in experimental group, blank control group and liposome control group was higher than that in control group. After transfection for 48 ~ 72 h, the Ishikawa cells of endometrial carcinoma in the experimental group increased faster than those in the other three groups. The Ishikawa cells of endometrial carcinoma in 72 ~ 96 h after transfection continued to increase and reached the logarithmic growth stage. The growth rate of Ishikawa cells in the experimental group was faster than that in the other three groups. The results of flow cytometry showed that the apoptosis rate of the experimental group was significantly lower than that of the other three groups after 3 days of transfection. The difference was statistically significant (P 0.05). The rate of apoptosis in the negative control group was significantly higher than that in the other three groups, and the difference was statistically significant. The results of RT-PCR showed that the expression of miR-183 in the experimental group was significantly lower than that in the other three groups. The difference was statistically significant (P 0.05). The expression of p53 mRNA in the experimental group was significantly lower than that in the other three groups. The expression of p53 protein in the experimental group was significantly higher than that in the other three groups. Conclusion miR-183 is expressed in Ishikawa cells of endometrial carcinoma. MiR-183 inhibitor can effectively inhibit the expression of miR-183 in endometrial carcinoma Ishikawa cells, promote cell proliferation and inhibit cell apoptosis. Upregulating the expression of p53 protein .miR-183 may inhibit the proliferation of endometrial carcinoma cells and promote apoptosis by inhibiting the expression of p53 protein, thus affecting the occurrence, development and prognosis of endometrial carcinoma.
【作者单位】：河北医科大学第三医院妇产科;伊犁哈萨克自治州友谊医院妇产科;
【分类号】：R737.33
【正文快照】： 子宫内膜癌是女性生殖系统常见恶性肿瘤之一,严重影响女性的健康和生命。随着分子生物学技术的发展,研究者们发现p53基因可能是子宫内膜癌发生、发展的重要基因,正常p53基因的缺失或突变可导致细胞无限生长,是肿瘤发生的一个早期事件。在人类恶性肿瘤患者中,至少50%的患者发生

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