本文關(guān)鍵詞：miR-183在子宮內(nèi)膜癌Ishikawa細(xì)胞中的表達及其對p53蛋白的調(diào)節(jié)作用　出處：《中國婦幼保健》2016年24期 　論文類型：期刊論文

  更多相關(guān)文章： 子宮內(nèi)膜癌 Ishikawa細(xì)胞 miR- p蛋白

【摘要】：目的探討miR-183在子宮內(nèi)膜癌Ishikawa細(xì)胞中的表達及其對p53蛋白表達和細(xì)胞增殖、凋亡的影響。方法采用LipofectaminerTM2000脂質(zhì)體將miR-183的特異性抑制劑反義寡核苷酸轉(zhuǎn)染到Ishikawa細(xì)胞(實驗組),不相關(guān)序列、脂質(zhì)體和未轉(zhuǎn)染組細(xì)胞分別為陰性對照組、脂質(zhì)體對照組和空白對照組。采用CCK-8法和流式細(xì)胞儀分別檢測各組子宮內(nèi)膜癌Ishikawa細(xì)胞的增殖和凋亡情況;采用RT-PCR檢測miR-183和p53 mRNA在各組子宮內(nèi)膜癌Ishikawa細(xì)胞中的表達水平;采用Western blot檢測p53蛋白在各組子宮內(nèi)膜癌Ishikawa細(xì)胞中的表達水平。結(jié)果 CCK-8檢測結(jié)果顯示:轉(zhuǎn)染0~24 h,實驗組、空白對照組、脂質(zhì)體對照組子宮內(nèi)膜癌Ishikawa細(xì)胞增速相似,陰性對照組子宮內(nèi)膜癌Ishikawa細(xì)胞增速略快;轉(zhuǎn)染24~48 h,實驗組、空白對照組和脂質(zhì)體對照組子宮內(nèi)膜癌Ishikawa細(xì)胞增速較快;轉(zhuǎn)染48~72 h,實驗組子宮內(nèi)膜癌Ishikawa細(xì)胞較其余3組增長速度快;轉(zhuǎn)染72~96 h,4組子宮內(nèi)膜癌Ishikawa細(xì)胞繼續(xù)保持增長狀態(tài),且達到對數(shù)生長期,實驗組子宮內(nèi)膜癌Ishikawa細(xì)胞較其余3組增速快。流式細(xì)胞學(xué)檢測結(jié)果顯示:轉(zhuǎn)染3 d后,實驗組較其余3組早期凋亡率明顯降低,差異有統(tǒng)計學(xué)意義(P0.05);陰性對照組較其余3組早期凋亡率明顯增高,差異有統(tǒng)計學(xué)意義(P0.05)。RT-PCR結(jié)果顯示:實驗組miR-183表達水平明顯低于其余3組,差異有統(tǒng)計學(xué)意義(P0.05);實驗組p53 mRNA表達水平明顯低于其余3組,差異有統(tǒng)計學(xué)意義(P0.05)。Western blot檢測結(jié)果顯示:實驗組p53蛋白表達水平明顯高于其余3組,差異有統(tǒng)計學(xué)意義(P0.05)。結(jié)論 miR-183在子宮內(nèi)膜癌Ishikawa細(xì)胞中有表達,miR-183抑制劑能有效抑制miR-183在子宮內(nèi)膜癌Ishikawa細(xì)胞中的表達,促進細(xì)胞增殖,抑制細(xì)胞凋亡,上調(diào)p53蛋白表達。miR-183可能通過抑制p53蛋白表達而抑制子宮內(nèi)膜癌細(xì)胞增殖,促進凋亡,從而影響子宮內(nèi)膜癌的發(fā)生、發(fā)展及預(yù)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.
【作者單位】： 河北醫(yī)科大學(xué)第三醫(yī)院婦產(chǎn)科;伊犁哈薩克自治州友誼醫(yī)院婦產(chǎn)科;
【分類號】：R737.33
【正文快照】： 子宮內(nèi)膜癌是女性生殖系統(tǒng)常見惡性腫瘤之一,嚴(yán)重影響女性的健康和生命。隨著分子生物學(xué)技術(shù)的發(fā)展,研究者們發(fā)現(xiàn)p53基因可能是子宮內(nèi)膜癌發(fā)生、發(fā)展的重要基因,正常p53基因的缺失或突變可導(dǎo)致細(xì)胞無限生長,是腫瘤發(fā)生的一個早期事件。在人類惡性腫瘤患者中,至少50%的患者發(fā)生

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