假基因PTENP1通过调控PTEN蛋白的表达影响胃癌进展的研究
发布时间：2019-03-19 16:01
【摘要】：背景:长链非编码RNA(long non-coding RNAs,lnc RNAs)是一类转录本长度超过200nt的RNA分子,它们缺乏蛋白编码的潜能。近年来,大量证据表明lnc RNAs能以RNA的形式在多种层面上(表观遗传调控、转录调控以及转录后调控等)调控基因的表达,从而在细胞增殖、细胞周期、分化和凋亡等多种生物学进程中发挥重要的作用。假基因转录得到的RNA也属于lnc RNA的一类,以前被认为没有生物学功能,然而最近一些假基因被证实在肿瘤的发生中发挥着关键的作用。PTENP1被发现在几种肿瘤细胞中发挥着抑癌作用,然而它在胃癌中的表达和生物学功能仍不清楚。目的:研究胃癌组织和细胞系中PTENP1的表达水平及其与临床病理资料的关系,并探索PTENP1在胃癌细胞中发挥的生物学功能及相关机制。方法:通过实时定量逆转录聚合酶链式反应(q RT-PCR)检测68例胃癌组织及4个胃癌细胞系中PTENP1的表达水平;通过去甲基化药物处理,研究PTENP1启动子异常甲基化与其表达的关系;分别转染p LJM-3’UTR或p LJM空质粒进入胃癌细胞系MGC-803及SGC-7901,通过CCK-8及平板克隆实验研究PTENP1对胃癌细胞增殖能力的影响,通过流式细胞分析及Hoechst staining实验研究PTENP1对细胞凋亡的影响,通过细胞划痕实验和transwell研究PTENP1对细胞迁移和侵袭能力的影响;通过q RT-PCR及Western blot分析质粒转染后PTEN的表达变化。结果:q RT-PCR检测结果显示PTENP1表达在胃癌组织和细胞系中显著下调,其表达下调可能部分与DNA超甲基化有关。而且,PTENP1的低表达与肿瘤的大小、胃癌病人临床分期、胃癌浸润深度及淋巴结转移有关。另外,结果显示PTENP1能够调节胃癌细胞的增殖、凋亡、迁移和侵袭。并且,PTENP1 3’UTR的表达升高能够提高PTEN蛋白的表达水平。结论:PTENP1在胃癌中表达显著下调,并能通过调节PTEN蛋白的表达发挥抑癌作用。
[Abstract]:Background: long-chain non-coding RNA (long non-coding RNAs,lnc RNAs) is a class of RNA molecules whose transcripts are longer than 200nt. They lack the potential of protein coding. In recent years, there has been a great deal of evidence that lnc RNAs can regulate the expression of genes at various levels (epigenetic regulation, transcriptional regulation and post-transcriptional regulation) in the form of RNA, thus, in cell proliferation and cell cycle, Differentiation and apoptosis play an important role in many biological processes. Pseudogene transcribed RNA also belongs to the lnc RNA family, which was previously thought to have no biological function. However, some pseudogenes have recently been proved to play a key role in tumorigenesis. PTENP1 has been found to play an anti-cancer role in several tumor cells, but its expression and biological function in gastric cancer are still unclear. Aim: to study the expression level of PTENP1 in gastric cancer tissues and cell lines and its relationship with clinicopathological data, and to explore the biological function and related mechanism of PTENP1 in gastric cancer cells. Methods: real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) was used to detect the expression of PTENP1 in 68 cases of gastric cancer tissues and 4 gastric cancer cell lines. The relationship between aberrant methylation of PTENP1 promoter and its expression was studied by demethylation drug treatment. The effects of PTENP1 on the proliferation of gastric cancer cell lines MGC-803 and SGC-7901, were studied by CCK-8 and plate cloning assay, respectively. The empty plasmids of p-LJM-3'UTR or p-LJM were transfected into gastric cancer cell line MGC-803 and SGC-7901, respectively. The effect of PTENP1 on cell apoptosis was studied by flow cytometry and Hoechst staining assay. The effects of PTENP1 on cell migration and invasion were studied by cell scratch test and transwell. The expression of PTEN was analyzed by Q-RT-PCR and Western blot. Results: the results of Q-RT-PCR showed that the expression of PTENP1 was significantly down-regulated in gastric cancer tissues and cell lines, and the down-regulation of the expression may be partly related to DNA hypermethylation. Furthermore, the low expression of PTENP1 was associated with tumor size, clinical stage, depth of invasion and lymph node metastasis. In addition, the results showed that PTENP1 can regulate the proliferation, apoptosis, migration and invasion of gastric cancer cells. In addition, the increased expression of PTENP1 3'UTR could increase the expression level of PTEN protein. Conclusion: the expression of PTENP1 is significantly down-regulated in gastric cancer, and can play an anti-cancer role by regulating the expression of PTEN protein.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.2
本文编号：2443671
[Abstract]:Background: long-chain non-coding RNA (long non-coding RNAs,lnc RNAs) is a class of RNA molecules whose transcripts are longer than 200nt. They lack the potential of protein coding. In recent years, there has been a great deal of evidence that lnc RNAs can regulate the expression of genes at various levels (epigenetic regulation, transcriptional regulation and post-transcriptional regulation) in the form of RNA, thus, in cell proliferation and cell cycle, Differentiation and apoptosis play an important role in many biological processes. Pseudogene transcribed RNA also belongs to the lnc RNA family, which was previously thought to have no biological function. However, some pseudogenes have recently been proved to play a key role in tumorigenesis. PTENP1 has been found to play an anti-cancer role in several tumor cells, but its expression and biological function in gastric cancer are still unclear. Aim: to study the expression level of PTENP1 in gastric cancer tissues and cell lines and its relationship with clinicopathological data, and to explore the biological function and related mechanism of PTENP1 in gastric cancer cells. Methods: real-time quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) was used to detect the expression of PTENP1 in 68 cases of gastric cancer tissues and 4 gastric cancer cell lines. The relationship between aberrant methylation of PTENP1 promoter and its expression was studied by demethylation drug treatment. The effects of PTENP1 on the proliferation of gastric cancer cell lines MGC-803 and SGC-7901, were studied by CCK-8 and plate cloning assay, respectively. The empty plasmids of p-LJM-3'UTR or p-LJM were transfected into gastric cancer cell line MGC-803 and SGC-7901, respectively. The effect of PTENP1 on cell apoptosis was studied by flow cytometry and Hoechst staining assay. The effects of PTENP1 on cell migration and invasion were studied by cell scratch test and transwell. The expression of PTEN was analyzed by Q-RT-PCR and Western blot. Results: the results of Q-RT-PCR showed that the expression of PTENP1 was significantly down-regulated in gastric cancer tissues and cell lines, and the down-regulation of the expression may be partly related to DNA hypermethylation. Furthermore, the low expression of PTENP1 was associated with tumor size, clinical stage, depth of invasion and lymph node metastasis. In addition, the results showed that PTENP1 can regulate the proliferation, apoptosis, migration and invasion of gastric cancer cells. In addition, the increased expression of PTENP1 3'UTR could increase the expression level of PTEN protein. Conclusion: the expression of PTENP1 is significantly down-regulated in gastric cancer, and can play an anti-cancer role by regulating the expression of PTEN protein.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R735.2
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