發(fā)布時(shí)間：2018-03-09 02:29

  本文選題：微小RNA　切入點(diǎn)：食管癌　出處：《中國(guó)現(xiàn)代醫(yī)學(xué)雜志》2017年13期 　論文類(lèi)型：期刊論文

【摘要】：目的探討micro RNA-196a(miR-196a)對(duì)食管癌預(yù)后的評(píng)估價(jià)值及其生物學(xué)行為的調(diào)控機(jī)制。方法收集120例行食管癌根治性手術(shù)切除的食管癌組織及癌旁組織,逆轉(zhuǎn)錄聚合酶鏈反應(yīng)(RT-PCR)檢測(cè)組織miR-196a表達(dá)水平,分析miR-196a表達(dá)與食管癌患者臨床資料的關(guān)系。對(duì)食管癌患者進(jìn)行隨訪(fǎng),記錄患者總生存期(OS)和無(wú)病生存期(DFS);以O(shè)S和DFS作為評(píng)價(jià)指標(biāo),采用單變量和多變量Cox比例風(fēng)險(xiǎn)模型評(píng)價(jià)患者預(yù)后的影響因素。分別采用miR-196a mimic、NC-mimic、miR-196a inhibitor、NC-inhibitor轉(zhuǎn)染食管癌TE1細(xì)胞,MTT實(shí)驗(yàn)檢測(cè)細(xì)胞增殖能力,Transwell實(shí)驗(yàn)檢測(cè)細(xì)胞侵襲能力,細(xì)胞劃痕實(shí)驗(yàn)檢測(cè)細(xì)胞遷移能力,Western blot檢測(cè)細(xì)胞ANXA1、NTN4、HMGA2、HOXB8蛋白表達(dá)。結(jié)果 miR-196a在食管癌組織中表達(dá)水平高于癌旁組織(P0.05);在TE1細(xì)胞中,miR-196a mimic組miR-196a表達(dá)水平高于NC-mimic組,miR-196a inhibitor組miR-196a表達(dá)水平低于NC-inhibitor組(P0.05),證實(shí)細(xì)胞轉(zhuǎn)染實(shí)驗(yàn)成功。將食管癌患者按照miR-196a表達(dá)分為miR-196a高表達(dá)組51例和低表達(dá)組69例,miR-196a表達(dá)與年齡、性別、分化程度、N分期、腫瘤位置等無(wú)關(guān)(P0.05),隨著T分期、TNM分期和腫瘤直徑增加,miR-196a高表達(dá)率升高(P0.05)。生存分析顯示,miR-196a高表達(dá)患者總生存期(P=0.015)和無(wú)病生存期(P=0.017)低于miR-196a低表達(dá)者;單因素和多因素分析顯示,T分期、miR-196a表達(dá)是影響患者總生存期和無(wú)病生存期的的獨(dú)立危險(xiǎn)因素(均P0.05)。MTT實(shí)驗(yàn)結(jié)果顯示,第48~120 h,miR-196a mimic組吸光度值高于NC-mimic組,miR-196a inhibitor組吸光度值低于NC-inhibitor組(P0.05);Transwell小室實(shí)驗(yàn)顯示,miR-196a mimic組穿膜細(xì)胞數(shù)量高于NC-mimic組,miR-196a inhibitor組穿膜細(xì)胞數(shù)量低于NC-inhibitor組(P0.05);細(xì)胞劃痕實(shí)驗(yàn)顯示,miR-196a mimic組細(xì)胞遷移距離高于NC-mimic組,miR-196a inhibitor組細(xì)胞遷移距離低于NC-inhibitor組(P0.05)。Western blot實(shí)驗(yàn)結(jié)果顯示,miR-196a mimic組、NC-mimic組、miR-196a inhibitor組、NC-inhibitor組HMGA2、HOXB8蛋白表達(dá)比較差異均無(wú)統(tǒng)計(jì)學(xué)意義(P0.05),miR-196a mimic組ANXA1、NTN4蛋白表達(dá)低于NC-mimic組,miR-196a inhibitor組ANXA1、NTN4蛋白表達(dá)高于NC-inhibitor組(P0.05)。結(jié)論 miR-196a能夠作為食管癌預(yù)后的預(yù)測(cè)指標(biāo)之一;miR-196a可能通過(guò)抑制ANXA1、NTN4基因的表達(dá)參與調(diào)控食管癌細(xì)胞的增殖、侵襲和遷移等生物學(xué)行為。
[Abstract]:Objective to investigate the value of micro RNA-196 miR-196a in evaluating the prognosis of esophageal carcinoma and the mechanism of its biological behavior. Methods 120 cases of esophageal carcinoma tissues and adjacent tissues were collected. Reverse transcriptase polymerase chain reaction (RT-PCR) was used to detect the expression of miR-196a and analyze the relationship between the expression of miR-196a and the clinical data of patients with esophageal carcinoma. Univariate and multivariate Cox proportional risk models were used to evaluate the prognostic factors of patients. The proliferation of esophageal carcinoma TE1 cells transfected with miR-196a mimicine NC-mimicum miR-196a inhibitor was measured by MTT assay and transwell assay was used to detect the invasion ability of esophageal cancer cells. Blot was used to detect the expression of ANXA1NTN4HMGA2OXB8 protein. Results the expression level of miR-196a in esophageal carcinoma tissue was higher than that in adjacent tissue P0.05A, and the miR-196a expression level in TE1 cells was higher than that in NC-mimic group. According to the expression of miR-196a, the patients with esophageal carcinoma were divided into miR-196a high expression group (51 cases) and low expression group (69 cases). Sex, degree of differentiation and N stage, tumor location were not related to P0.05, but with the increase of T stage and tumor diameter, the high expression rate of miR-196a increased (P 0.05). Survival analysis showed that the total survival time (P0.015) and disease-free survival time (P0.017) of patients with high expression of miR-196a were lower than those of patients with low expression of miR-196a. Univariate and multivariate analysis showed that the expression of miR-196a in T stage was an independent risk factor for the overall survival and disease-free survival of patients (all P0.05. The absorbance of miR-196a mimic group was higher than that of NC-mimic group inhibitor group was lower than that of NC-inhibitor group (P0.05) Transwell chamber experiment showed that the number of perforating cells in mimic group was higher than that in NC-mimic group (P 0.05), and the cell scratch test showed that the number of perforating cells in NC-mimic group was lower than that in NC-inhibitor group (P0.05), and the cell scratch test showed that the number of perforating cells in mimic group was lower than that in NC-inhibitor group (P0.05). Cell migration distance was higher than that in NC-mimic group (P 0.05). Western blot assay showed that the expression of HMGA2HXB8 protein in NC-mimic group of mimic group was lower than that in mimic group of NC-mimic group. There was no significant difference in the expression of HMGA2HOXB8 protein in NC-inhibitor group. There was no significant difference in the expression of ANXA1nTN4 protein in mimic group of NC-mimic group compared with that in mimic group of NC-mimic group (P0.05miR-196a mimic group). The expression of ANXA1nTN4 protein in mimic group was lower than that in mimic group of NC-mimic group. Conclusion the expression of ANXA1nTN4 protein is higher than that of NC-inhibitor group P0.05.Conclusion miR-196a may play an important role in regulating the proliferation of esophageal cancer cells by inhibiting the expression of ANXA1NTN4 gene. Biological behaviors such as invasion and migration.
【作者單位】： 河南科技大學(xué)第一附屬醫(yī)院腫瘤外科二病區(qū);
【分類(lèi)號(hào)】：R735.1
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本文編號(hào)：1586622