本文選題：微小核糖核酸--p + 宮頸癌�。� 參考：《中國腫瘤生物治療雜志》2017年07期

【摘要】：目的:觀察微小核糖核酸-134-5p(mi R-134-5p)轉(zhuǎn)染對宮頸癌細(xì)胞增殖和凋亡的影響,驗(yàn)證其可能的分子機(jī)制。方法:收集湖北醫(yī)藥學(xué)院附屬人民醫(yī)院腫瘤中心2016年5月至8月收治的8名宮頸癌患者腫瘤組織和相應(yīng)癌旁組織。利用lipofectamine 2000將mi R-134-5p mimics轉(zhuǎn)染至宮頸癌Hela和Si Ha細(xì)胞。采用MTT法和集落形成實(shí)驗(yàn)檢測細(xì)胞增殖活性;流式細(xì)胞術(shù)(FCM)檢測細(xì)胞周期和細(xì)胞凋亡;q RT-PCR檢測宮頸癌組織和細(xì)胞mi R-134-5p m RNA表達(dá)以及宮頸癌細(xì)胞EGFR m RNA表達(dá);Western blotting檢測宮頸癌細(xì)胞EGFR信號通路相關(guān)蛋白的表達(dá)。結(jié)果:宮頸癌組織mi R-134-5p m RNA表達(dá)顯著低于癌旁組織(P0.01)。和轉(zhuǎn)染mi R-NC的Hela和Si Ha細(xì)胞比較,轉(zhuǎn)染mi R-134-5pmimics的宮頸癌Hela和Si Ha細(xì)胞mi R-134-5p m RNA表達(dá)顯著升高;細(xì)胞增殖能力顯著降低(轉(zhuǎn)染第5天,Hela細(xì)胞:1.06±0.13 vs 1.32±0.07;Si Ha細(xì)胞:1.12±0.10 vs 1.42±0.12,均P0.05);形成的集落數(shù)減少;G0/G1期細(xì)胞比例顯著上升,S期和G2/M期細(xì)胞比例顯著下降;細(xì)胞凋亡率顯著增加[Hela細(xì)胞:(26.53±13.48)%vs(3.25±1.74)%;Si Ha細(xì)胞:(30.49±12.04)%vs(5.10±2.86)%,均P0.05];EGFR m RNA和EGFR蛋白表達(dá)顯著下調(diào),其中EGFR m RNA,Hela細(xì)胞下調(diào)58%(P0.01),Si Ha細(xì)胞下調(diào)41%(P0.05);EGFR下游靶蛋白p-AKT、p-ERK1/2和Cyclin D1蛋白及p EGFR蛋白表達(dá)顯著下調(diào)。結(jié)論:mi R-134-5p可顯著抑制宮頸癌細(xì)胞增殖并促進(jìn)細(xì)胞凋亡,其可能的分子機(jī)制是通過抑制EGFR基因的表達(dá),抑制EGFR通路的活化。
[Abstract]:Aim: to investigate the effect of RNC-134-5pmmi R-134-5p) transfection on the proliferation and apoptosis of cervical cancer cells, and to verify its possible molecular mechanism. Methods: the tumor tissues and adjacent tissues of 8 patients with cervical cancer were collected from the Cancer Center of the people's Hospital affiliated to Hubei Medical College from May to August 2016. Mi R-134-5p mimics was transfected into cervical cancer Hela and SIHA cells by lipofectamine 2000. Cell proliferation activity was detected by MTT assay and colony forming assay. Flow cytometry (FCM) was used to detect the expression of mi R-134-5p m RNA in cervical carcinoma tissues and cells, and the expression of EGFR signal pathway related protein in cervical cancer cells by EGFR m RNA blotting. Results: the expression of mi R-134-5p m RNA in cervical carcinoma was significantly lower than that in paracancerous tissue (P 0.01). Compared with Hela and SIHA cells transfected with mi R-NC, the expression of mi R-134-5p m RNA in Hela and SIHA cells transfected with mi R-134-5pmimics was significantly increased. On the 5th day after transfection, the proliferation of Hela cells was significantly decreased (1.06 鹵0.13 vs 1.32 鹵0.07Si Ha: 1.12 鹵0.10 vs 1.42 鹵0.12, all P 0.05), and the number of colony formation decreased significantly in G _ 0 / G _ 1 phase and G _ 2 / M phase. The expression of EGFR-m RNA and EGFR protein were significantly down-regulated in Hela cells. EGFR m RNA-Hela cells down-regulated the down-regulation of the downstream target proteins of 41P0.05EGFR, p-AKTnp-ERK1and Cyclin D1 protein and pEGFR protein in Hela cells (P < 0.05). The apoptosis rate was significantly increased in Hela cells (P < 0.01 鹵1.74), and the expression of EGFR-m RNA and EGFR protein was significantly down-regulated in Hela cell line (P0.05EGFR downstream target protein p-AKTFP-ER12 and Cyclin D1 protein and p EGFR protein). The expression of p-AKTP-ERK1.2 and Cyclin D1 protein and p EGFR protein were down-regulated by EGFR m RNA-Hela cells. Conclusion: the cell proliferation and apoptosis of cervical cancer cells were significantly inhibited by 1: mi R-134-5p. The possible molecular mechanism was to inhibit the activation of EGFR pathway by inhibiting the expression of EGFR gene.
【作者單位】： 湖北醫(yī)藥學(xué)院附屬人民醫(yī)院腫瘤中心;湖北醫(yī)藥學(xué)院附屬人民醫(yī)院中藥藥理實(shí)驗(yàn)室;
【基金】：湖北省教育廳科學(xué)技術(shù)研究資助項(xiàng)目(No.B2016139) 湖北省高校優(yōu)秀中青年創(chuàng)新基金資助項(xiàng)目(No.T201510)~~
【分類號】：R737.33

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