發(fā)布時(shí)間：2017-12-31 07:31

  本文關(guān)鍵詞：miR-29a-3p對(duì)胃癌腹膜轉(zhuǎn)移的影響及其機(jī)制研究　出處：《寧夏醫(yī)科大學(xué)》2016年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： miR-29a-3p 胃癌 增殖轉(zhuǎn)移 HAS3

【摘要】：背景:胃癌是最常見的惡性腫瘤之一,其發(fā)病率在世界范圍惡性腫瘤排第四位,死亡率居第二位,在我國(guó),每年新發(fā)病例約為30萬(wàn),嚴(yán)重威脅著國(guó)人健康,而轉(zhuǎn)移是胃癌患者最常見的致死原因,其發(fā)生的分子機(jī)制一直是腫瘤研究領(lǐng)域的熱點(diǎn)。微小RNA(micro RNAs,mi RNAs)是一大類內(nèi)源性的、在進(jìn)化過(guò)程中高度保守的單鏈非編碼小RNA,參與了人體多種生命活動(dòng)的調(diào)控,如細(xì)胞的增殖、分化、凋亡等。mi RNA的表達(dá)失調(diào)可導(dǎo)致多種疾病的發(fā)生,并且與多種惡性腫瘤的發(fā)生、發(fā)展有著極為密切的關(guān)系。近年來(lái)許多研究表明,在胃癌中也存在mi RNA表達(dá)失調(diào)的現(xiàn)象,說(shuō)明mi RNA也參與了胃癌的發(fā)生。本實(shí)驗(yàn)將在前期的基礎(chǔ)上,對(duì)mi R-29a-3p在胃癌細(xì)胞系的表達(dá)水平、生物學(xué)功能及可能的作用機(jī)制進(jìn)行初步的研究和探討。目的:探索hsa-mi R-29a-3p(mi R-29a)對(duì)胃癌腹膜轉(zhuǎn)移的影響并初步研究其作用機(jī)制。方法:1.利用q RT-PCR驗(yàn)證mi R-29a-3p在三對(duì)胃癌高低轉(zhuǎn)移細(xì)胞系GC9811-P/GC9811,MKN28-M/MKN-28NM,SGC7901-M/SGC7901-NM中的表達(dá)情況。2.構(gòu)建過(guò)表達(dá)及抑制mi R-29a-3p的慢病毒表達(dá)載體,將過(guò)表達(dá)mi R-29a-3p的慢病毒及其無(wú)關(guān)陰性對(duì)照轉(zhuǎn)染入胃癌腹膜高轉(zhuǎn)移細(xì)胞系GC9811-P中,相反,將抑制mi R-29a-3p表達(dá)的慢病毒及其無(wú)關(guān)陰性對(duì)照轉(zhuǎn)入親本細(xì)胞系GC9811中。3.用MTT比色實(shí)驗(yàn)、平板克隆形成實(shí)驗(yàn)、Transwell、劃痕實(shí)驗(yàn)及流式細(xì)胞學(xué)技術(shù)分別觀察mi R-29a-3p對(duì)胃癌細(xì)胞的增殖、轉(zhuǎn)移及凋亡能力的影響。4.采用生物信息學(xué)的方法預(yù)測(cè)mi R-29a-3p的靶基因,查閱文獻(xiàn)選出與轉(zhuǎn)移相關(guān)的基因并采用q RT-PCR及western blot驗(yàn)證與mi R-29a-3p的關(guān)系。5.轉(zhuǎn)染si RNA及空載體沉默基因表達(dá),進(jìn)一步行功能試驗(yàn)驗(yàn)證其功能及是否為mi R-29a-3p的靶基因。結(jié)果:1.mi R-29a-3p在胃癌高轉(zhuǎn)移細(xì)胞系(GC9811-P、MKN28-M、SGC7901-M)中高表達(dá)。2.成功構(gòu)建高低表達(dá)mi R-29a-3p的細(xì)胞系,q RT-PCR驗(yàn)證轉(zhuǎn)染成功。3.功能試驗(yàn)結(jié)果表明,過(guò)表達(dá)mi R-29a-3p可以抑制細(xì)胞的增殖及轉(zhuǎn)移能力而促進(jìn)細(xì)胞的凋亡能力;相反,抑制mi R-29a-3p的表達(dá)可以促進(jìn)細(xì)胞的增殖及轉(zhuǎn)移能力而抑制細(xì)胞的凋亡能力。4.生物信息學(xué)預(yù)測(cè)提示HAS3可能為mi R-29a-3p的潛在靶基因之一,q RT-PCR結(jié)果顯示,在上調(diào)mi R-29a-3p的細(xì)胞系中,HAS3表達(dá)有降低的趨勢(shì)但無(wú)統(tǒng)計(jì)學(xué)意義;在下調(diào)mi R-29a-3p的細(xì)胞系中,HAS3表達(dá)有升高的趨勢(shì)但無(wú)統(tǒng)計(jì)學(xué)意義。Western blot結(jié)果顯示,上調(diào)mi R-29a-3p的表達(dá),HAS3表達(dá)下調(diào),下調(diào)mi R-29a-3p的表達(dá),HAS3表達(dá)上調(diào)。5.沉默HAS3的表達(dá),胃癌細(xì)胞增殖及遷移能力減弱,說(shuō)明HAS3促進(jìn)胃癌細(xì)胞的增殖與轉(zhuǎn)移。在高表達(dá)mi R-29a-3p的細(xì)胞系中沉默HAS3的表達(dá)后,細(xì)胞的增殖及轉(zhuǎn)移能力減弱,與過(guò)表達(dá)mi R-29a-3p有類似的效果,說(shuō)明mi R-29a-3p可負(fù)性調(diào)控HAS3,進(jìn)一步證實(shí)HAS3可為mi R-29a-3p的潛在靶基因之一。結(jié)論:mi R-29a-3p抑制胃癌細(xì)胞的增殖與轉(zhuǎn)移而促進(jìn)凋亡,并可能通過(guò)其下游靶分子之一HAS3發(fā)揮其作用。
[Abstract]:Background: gastric cancer is one of the most common malignant tumors, the incidence of cancer ranked 4th in the world, the death rate ranked second. In China, the annual new cases are about 300,000, which seriously threaten the health of people. Metastasis is the most common cause of death in patients with gastric cancer, and its molecular mechanism has been a hot spot in the field of cancer research. Small RNA(micro RNAs. Mi RNAss are a large class of endogenous, highly conserved, single-stranded, non-coding RNAs that participate in the regulation of a variety of human life activities, such as cell proliferation and differentiation. The expression imbalance of .mi RNA, such as apoptosis, can lead to the occurrence of many diseases, and is closely related to the occurrence and development of many malignant tumors. There is also an imbalance in the expression of mi RNA in gastric cancer, indicating that mi RNA is also involved in the development of gastric cancer. The expression level of mi R-29a-3p in gastric cancer cell line. Biological function and possible mechanism of action were preliminarily studied and discussed. Objective: to explore hsa-mi R-29a-3pmmi R-29a. Effect on peritoneal metastasis of gastric cancer and preliminary study of its mechanism. Methods: 1. Using Q RT-PCR to verify mi. R-29a-3p was detected in three pairs of gastric cancer cell lines GC9811-P/GC9811. MKN28-M/MKN-28NM. The expression of SGC7901-M/SGC7901-NM. 2. Construct the expression vector of lentivirus which overexpressed and inhibited mi R-29a-3p. The overexpression of mi R-29a-3p lentivirus and its unrelated negative control were transfected into the peritoneal high metastasis cell line GC9811-P of gastric cancer. The lentivirus which inhibited the expression of mi R-29a-3p and its unrelated negative control were transferred into the parental cell line GC9811. 3. MTT colorimetric assay and plate clone formation test were used. The proliferation of gastric cancer cells induced by miR-29a-3p was observed by flow cytometry and scratch test. Effects of Metastasis and apoptosis. Bioinformatics was used to predict the target gene of mi R-29a-3p. Search the literature to select genes related to metastasis and verify the relationship with mi R-29a-3p by Q RT-PCR and western blot .5.transfect si. RNA and empty vector silenced gene expression. Further functional tests were performed to verify its function and whether it was the target gene of mi R-29a-3p. Results: 1. Mi R-29a-3p was detected in gastric cancer cell line GC9811-P. High expression of MKN28-MGC7901-M2.The cell line with high and low expression of miR-29a-3p was successfully constructed. The results of functional test showed that overexpression of mi R-29a-3p could inhibit cell proliferation and metastasis and promote cell apoptosis. On the contrary. Inhibition of the expression of mi R-29a-3p could promote cell proliferation and metastasis and inhibit cell apoptosis. 4. Bioinformatics prediction suggested that HAS3 might be mi. One of the potential target genes of R-29a-3p. The results of Q RT-PCR showed that the expression of HAS3 decreased in the cell line of miR-29a-3p, but had no statistical significance. The down-regulation of the expression of HAS3 in the cell line of miR-29a-3p showed a tendency to increase, but no statistical significance. Western blot results showed. Up regulated the expression of miR-29a-3p and down-regulated the expression of miR-29a-3p and up-regulated the expression of HAS3. Silenced the expression of HAS3. The ability of proliferation and migration of gastric cancer cells was decreased, which indicated that HAS3 promoted proliferation and metastasis of gastric cancer cells. After silencing the expression of HAS3 in the cell lines with high expression of miR-29a-3p. The ability of cell proliferation and metastasis was decreased, which was similar to that of overexpression of mi R-29a-3p, indicating that mi R-29a-3p could negatively regulate HAS3. It is further confirmed that HAS3 may be one of the potential target genes of mi R-29a-3p. Conclusion: MiR-29a-3p inhibits the proliferation and metastasis of gastric cancer cells and promotes apoptosis. And it may play its role through one of its downstream target molecules, HAS3.
【學(xué)位授予單位】：寧夏醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2016
【分類號(hào)】：R735.2

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 戴冬秋;胃癌腹膜轉(zhuǎn)移研究之策略[J];世界華人消化雜志;2005年09期

2 丁松濤;張忠濤;;胃癌腹膜轉(zhuǎn)移的研究現(xiàn)狀和進(jìn)展[J];國(guó)際外科學(xué)雜志;2006年06期

3 李正榮;詹文華;;胃癌腹膜轉(zhuǎn)移的治療進(jìn)展[J];中華胃腸外科雜志;2006年05期

4 聶秉宇;姜鑫;白秀峰;張杰;于年華;程柱清;;老年胃癌腹膜轉(zhuǎn)移的研究進(jìn)展[J];中國(guó)老年學(xué)雜志;2007年16期

5 張賢坤;朱萬(wàn)坤;韓曉鵬;蘇玉林;李文惠;劉宏斌;;胃癌腹膜轉(zhuǎn)移的診斷及防治[J];中國(guó)腫瘤臨床;2008年15期

6 趙宏偉;劉宏斌;李文惠;;胃癌腹膜轉(zhuǎn)移機(jī)理研究現(xiàn)狀[J];現(xiàn)代腫瘤醫(yī)學(xué);2009年05期

7 劉移忠;;胃癌腹膜轉(zhuǎn)移的螺旋CT診斷[J];中國(guó)當(dāng)代醫(yī)藥;2011年17期

8 張海峰;張傳珉;;胃癌腹膜轉(zhuǎn)移診治的研究進(jìn)展[J];中國(guó)癌癥防治雜志;2011年02期

9 趙璐;趙愛光;;胃癌腹膜轉(zhuǎn)移治療的研究進(jìn)展[J];世界華人消化雜志;2011年26期

10 趙璐;趙愛光;;胃癌腹膜轉(zhuǎn)移的實(shí)驗(yàn)診斷進(jìn)展[J];中國(guó)實(shí)驗(yàn)診斷學(xué);2012年10期

相關(guān)會(huì)議論文 前2條

1 華子辰;嚴(yán)超;燕敏;朱正綱;;血清CA125在胃癌腹膜轉(zhuǎn)移中的臨床價(jià)值[A];第9屆全國(guó)胃癌學(xué)術(shù)會(huì)議暨第二屆陽(yáng)光長(zhǎng)城腫瘤學(xué)術(shù)會(huì)議論文匯編[C];2014年

2 邵麗華;劉少平;鐘燕軍;方敏;劉洋;李雁;;新型阿霉素前體藥PADM治療胃癌腹膜轉(zhuǎn)移癌的實(shí)驗(yàn)研究[A];中華醫(yī)學(xué)會(huì)腫瘤學(xué)分會(huì)第七屆全國(guó)中青年腫瘤學(xué)術(shù)會(huì)議——中華醫(yī)學(xué)會(huì)腫瘤學(xué)分會(huì)“中華腫瘤 明日之星”大型評(píng)選活動(dòng)暨中青年委員全國(guó)遴選論文匯編[C];2011年

相關(guān)重要報(bào)紙文章 前1條

1 記者 高翔;胃癌腹膜轉(zhuǎn)移治療有突破[N];健康報(bào);2011年

相關(guān)博士學(xué)位論文 前6條

1 華子辰;血清CA125在胃癌腹膜轉(zhuǎn)移中的臨床價(jià)值[D];上海交通大學(xué);2014年

2 趙丹懿;胃癌腹膜轉(zhuǎn)移相關(guān)標(biāo)志物的初步篩選及其功能研究[D];中國(guó)醫(yī)科大學(xué);2007年

3 張磊;胃癌腹膜轉(zhuǎn)移相關(guān)蛋白Myosin-9促進(jìn)胃癌的侵襲和轉(zhuǎn)移[D];南方醫(yī)科大學(xué);2014年

4 趙群;脾虛型胃癌腹膜轉(zhuǎn)移的生物學(xué)特性以及中西醫(yī)結(jié)合治療的基礎(chǔ)與臨床研究[D];河北醫(yī)科大學(xué);2003年

5 唐利;新型阿霉素前體藥PDOX聯(lián)合細(xì)胞減滅術(shù)加腹腔熱灌注化療治療胃癌腹膜轉(zhuǎn)移癌的實(shí)驗(yàn)研究[D];武漢大學(xué);2014年

6 邵麗華;新型阿霉素前體藥PDOX藥效及安全性實(shí)驗(yàn)研究[D];武漢大學(xué);2013年

相關(guān)碩士學(xué)位論文 前10條

1 張海峰;胃癌腹膜轉(zhuǎn)移的臨床特點(diǎn)及手術(shù)治療結(jié)果分析[D];廣西醫(yī)科大學(xué);2012年

2 楊顯金;腹膜間皮細(xì)胞間質(zhì)化在胃癌腹膜轉(zhuǎn)移中的研究[D];川北醫(yī)學(xué)院;2015年

3 辛瑞娟;miRNA-214在胃癌腹膜轉(zhuǎn)移中的作用及機(jī)制研究[D];寧夏醫(yī)科大學(xué);2015年

4 酒夢(mèng)娜;miR-29a-3p對(duì)胃癌腹膜轉(zhuǎn)移的影響及其機(jī)制研究[D];寧夏醫(yī)科大學(xué);2016年

5 惠亮亮;血紅素加氧酶-1在胃癌腹膜轉(zhuǎn)移中的作用及其相關(guān)機(jī)制研究[D];寧夏醫(yī)科大學(xué);2010年

6 曾若蘭;溶血磷脂酸介導(dǎo)胃癌腹膜轉(zhuǎn)移的研究[D];中南大學(xué);2014年

7 高國(guó)鋒;Peroxiredoxin-1在胃癌腹膜轉(zhuǎn)移中的作用及相關(guān)機(jī)制的研究[D];山西醫(yī)科大學(xué);2014年

8 劉克波;Eph-B_2受體在胃癌組織中的表達(dá)及其與胃癌腹膜轉(zhuǎn)移的關(guān)系[D];中南大學(xué);2010年

9 劉選文;黏附分子P-選擇素在胃癌腹膜轉(zhuǎn)移中的表達(dá)及臨床意義[D];大連醫(yī)科大學(xué);2011年

10 韓嘯;趨化因子受體CXCR7，CCR5在胃癌腹膜轉(zhuǎn)移中的表達(dá)及意義[D];大連醫(yī)科大學(xué);2012年

，

本文編號(hào)：1358788