發(fā)布時(shí)間：2018-01-31 13:25

  本文關(guān)鍵詞： GPRCA 前列腺癌 上皮間質(zhì)轉(zhuǎn)化　出處：《中國(guó)生物工程雜志》2017年03期 　論文類型：期刊論文

【摘要】：目的:構(gòu)建GPRC6A基因過表達(dá)的前列腺癌LncapC4-2細(xì)胞株,檢測(cè)細(xì)胞遷移和侵襲能力改變及EMT相關(guān)基因表達(dá),為研究GPRC6A介導(dǎo)的EMT在前列腺癌發(fā)生發(fā)展過程中的作用奠定基礎(chǔ)。方法:構(gòu)建過表達(dá)GPRC6A的慢病毒載體pCDH-GPRC6A,人胚腎293T細(xì)胞包被病毒并感染Lncap C4-2細(xì)胞株,Puromycin篩選獲得穩(wěn)定過表達(dá)GPRC6A的細(xì)胞株。RT-PCR和Western blot驗(yàn)證細(xì)胞GPRC6A的過表達(dá)情況。劃痕和transwell實(shí)驗(yàn)檢測(cè)細(xì)胞遷移和侵襲能力,qPCR檢測(cè)EMT相關(guān)基因表達(dá)。結(jié)果:成功構(gòu)建GPRC6A表達(dá)的慢病毒載體,RT-PCR和Western blot檢測(cè)LncapC4-2 GPRC6A過表達(dá)細(xì)胞株中GPRC6A的mRNA和蛋白質(zhì)表達(dá)增高,與對(duì)照相比較,在過表達(dá)GPRC6A的LncapC4-2細(xì)胞中,細(xì)胞遷移和侵襲能力增強(qiáng),并且EMT相關(guān)基因E-cadherin表達(dá)降低而SNAIL表達(dá)增高。結(jié)論:成功構(gòu)建過表達(dá)的GPRC6A的前列腺癌細(xì)胞株,腫瘤細(xì)胞中的GPRC6A可能通過增強(qiáng)細(xì)胞遷移和侵襲能力并促進(jìn)細(xì)胞EMT而參與前列腺癌發(fā)生發(fā)展過程。
[Abstract]:Objective: to construct a prostate cancer LncapC4-2 cell line with overexpression of GPRC6A gene and to detect the change of migration and invasion ability and the expression of EMT related genes. To study the role of EMT mediated by GPRC6A in the development of prostate cancer. Methods: a lentivirus vector pCDH-GPRC6A was constructed to express GPRC6A. Human embryonic kidney 293T cells were coated with virus and infected with Lncap C4-2 cell line. Puromycin screening of cells with stable overexpression of GPRC6A. RT-PCR and Western. Blot was used to verify the overexpression of GPRC6A. Scratch and transwell assay were used to detect cell migration and invasion. QPCR was used to detect the expression of EMT related genes. Results: the lentivirus vector expressing GPRC6A was successfully constructed. RT-PCR and Western blot were used to detect the increase of mRNA and protein expression of GPRC6A in LncapC4-2 GPRC6A overexpression cell lines. Compared with the control group, the migration and invasion of LncapC4-2 cells with overexpression of GPRC6A were enhanced. The expression of E-cadherin was decreased and the expression of SNAIL was increased. Conclusion: the prostate cancer cell line with overexpression of GPRC6A was successfully constructed. GPRC6A in tumor cells may be involved in the development of prostate cancer by enhancing cell migration and invasion and promoting cellular EMT.
【作者單位】： 山西醫(yī)科大學(xué)基礎(chǔ)醫(yī)學(xué)院;北京協(xié)和醫(yī)學(xué)院研究生院;
【基金】：國(guó)家青年科學(xué)基金(81302220) 山西省青年科學(xué)基金(2014021037-1) 山西醫(yī)科大學(xué)博士啟動(dòng)基金(B03201204)資助項(xiàng)目
【分類號(hào)】：R737.25
【正文快照】： 近年來,隨著我國(guó)老齡化人口的增長(zhǎng),前列腺癌正成為嚴(yán)重影響我國(guó)男性健康的泌尿系惡性腫瘤[1-2]。前列腺癌好發(fā)骨轉(zhuǎn)移,加重病情的發(fā)展[3]。有必要尋找前列腺癌骨轉(zhuǎn)移的標(biāo)志物,為前列腺癌骨轉(zhuǎn)移有效防治提供新思路和藥物作用的新靶點(diǎn)。GPRC6A是2004年發(fā)現(xiàn)的G蛋白耦聯(lián)受體C家族成

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1 劉銘;中國(guó)北方人群前列腺癌風(fēng)險(xiǎn)位點(diǎn)識(shí)別和GPRC6A基因功能的研究[D];北京協(xié)和醫(yī)學(xué)院;2012年

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