發(fā)布時間：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基因過表達的前列腺癌LncapC4-2細胞株,檢測細胞遷移和侵襲能力改變及EMT相關(guān)基因表達,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GPRC6A的慢病毒載體pCDH-GPRC6A,人胚腎293T細胞包被病毒并感染Lncap C4-2細胞株,Puromycin篩選獲得穩(wěn)定過表達GPRC6A的細胞株。RT-PCR和Western blot驗證細胞GPRC6A的過表達情況。劃痕和transwell實驗檢測細胞遷移和侵襲能力,qPCR檢測EMT相關(guān)基因表達。結(jié)果:成功構(gòu)建GPRC6A表達的慢病毒載體,RT-PCR和Western blot檢測LncapC4-2 GPRC6A過表達細胞株中GPRC6A的mRNA和蛋白質(zhì)表達增高,與對照相比較,在過表達GPRC6A的LncapC4-2細胞中,細胞遷移和侵襲能力增強,并且EMT相關(guān)基因E-cadherin表達降低而SNAIL表達增高。結(jié)論:成功構(gòu)建過表達的GPRC6A的前列腺癌細胞株,腫瘤細胞中的GPRC6A可能通過增強細胞遷移和侵襲能力并促進細胞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)科大學基礎(chǔ)醫(yī)學院;北京協(xié)和醫(yī)學院研究生院;
【基金】：國家青年科學基金(81302220) 山西省青年科學基金(2014021037-1) 山西醫(yī)科大學博士啟動基金(B03201204)資助項目
【分類號】：R737.25
【正文快照】： 近年來,隨著我國老齡化人口的增長,前列腺癌正成為嚴重影響我國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。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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