本文關(guān)鍵詞：人剪切修復(fù)基因XPD抑制肝癌HepG2細(xì)胞增殖遷移及對(duì)自噬水平調(diào)控的實(shí)驗(yàn)研究　出處：《南昌大學(xué)》2015年碩士論文　論文類(lèi)型：學(xué)位論文

  更多相關(guān)文章： 肝癌 XPD 增殖 凋亡 遷移 自噬

【摘要】：目的:探究上調(diào)肝癌細(xì)胞株HepG2中XPD基因的表達(dá)后對(duì)腫瘤細(xì)胞增殖凋亡、遷移以及自噬水平變化的影響。方法:采用脂質(zhì)體作為載體介質(zhì)將重組質(zhì)粒pEGFP-N2/XPD和空載質(zhì)粒pEGFP-N2瞬時(shí)轉(zhuǎn)染入人肝癌細(xì)胞株HepG2細(xì)胞內(nèi),實(shí)驗(yàn)分為四組1)空白對(duì)照組、2)脂質(zhì)體組、3)pEGFP-N2組、4)pEGFP-N2/XPD組,使用熒光顯微鏡觀察綠色熒光表達(dá)情況;MTT法觀察腫瘤細(xì)胞的增殖能力;流式細(xì)胞儀檢測(cè)腫瘤細(xì)胞凋亡情況;體外平板克隆形成實(shí)驗(yàn)觀察腫瘤細(xì)胞體外克隆集落形成情況;劃痕愈合實(shí)驗(yàn)及Transwell實(shí)驗(yàn)檢測(cè)腫瘤細(xì)胞遷移能力的變化。Western blotting檢測(cè)HepG2細(xì)胞XPD及自噬相關(guān)蛋白LC3、P62表達(dá)量的變化。結(jié)果:熒光顯微鏡觀察轉(zhuǎn)染空載質(zhì)粒pEGFP-N2和重組質(zhì)粒pEGFPN2/XPD的細(xì)胞可見(jiàn)綠色熒光,轉(zhuǎn)染有效。上調(diào)XPD的表達(dá)后,結(jié)果顯示與對(duì)照組比較,實(shí)驗(yàn)組細(xì)胞增殖能力減弱(p0.01),細(xì)胞凋亡率增加(p0.01),體外克隆形成數(shù)量減少(p0.01),遷移能力減弱(p0.05),并且上調(diào)LC3蛋白表達(dá)(p0.01),而隨之P62的表達(dá)水平減少(p0.01)。結(jié)論:過(guò)表達(dá)XPD基因能顯著抑制肝癌細(xì)胞增殖促進(jìn)其凋亡,抑制體外克隆形成,并且降低腫瘤細(xì)胞的遷移能力,同時(shí)調(diào)控肝癌細(xì)胞自噬水平的活化。
[Abstract]:Objective: to investigate the proliferation and apoptosis of hepatoma cells after up-regulating the expression of XPD gene in hepatoma cell line HepG2. Effects of migration and changes in autophagy levels. Methods:. The recombinant plasmid pEGFP-N2/XPD and the empty plasmid pEGFP-N2 were transiently transfected into human hepatoma cell line HepG2 cells using liposome as carrier medium. The experiment was divided into four groups (1) blank control group (2)) liposome group (pEGFP-N2) and pEGFP-N _ 2 / XPD group. The expression of green fluorescence was observed by fluorescence microscope. The proliferative ability of tumor cells was observed by MTT method. Apoptosis of tumor cells was detected by flow cytometry. The colony formation of tumor cells in vitro was observed by plate clone formation in vitro. Effect of scratch healing test and Transwell assay on the migration ability of tumor cells. XPD and autophagy associated protein LC3 in HepG2 cells were detected by blotting. Results: green fluorescence was observed in the cells transfected with empty plasmid pEGFP-N2 and recombinant plasmid pEGFPN2/XPD. After up-regulating the expression of XPD, the results showed that compared with the control group, the proliferation ability of the experimental group decreased p0.01a, and the apoptosis rate increased (p0.01). The number of clone formation in vitro decreased p0.01a, the migration capacity decreased, and the expression of LC3 protein was upregulated (p0.01). Conclusion: overexpression of XPD gene can significantly inhibit the proliferation of hepatoma cells and promote its apoptosis, and inhibit the formation of in vitro cloning. It also reduces the migration ability of tumor cells and regulates the activation of autophagy of hepatoma cells.
【學(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類(lèi)號(hào)】：R735.7

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 Javier Tejeda-Maldonado;Ignacio García-Juárez;Jonathan Aguirre-Valadez;Adrián González-Aguirre;Mario Vilatobá-Chapa;Alejandra Armengol-Alonso;Francisco Escobar-Penagos;Aldo Torre;Juan Francisco Sánchez-ávila;Diego Luis Carrillo-Pérez;;Diagnosis and treatment of hepatocellular carcinoma: An update[J];World Journal of Hepatology;2015年03期

2 Maria-Aggeliki Kalogeridi;Anna Zygogianni;George Kyrgias;John Kouvaris;Sofia Chatziioannou;Nikolaos Kelekis;Vassilis Kouloulias;;Role of radiotherapy in the management of hepatocellular carcinoma:A systematic review[J];World Journal of Hepatology;2015年01期

3 張慶燕;趙慧;羅文;張吉翔;;人剪切修復(fù)基因XPD肝癌對(duì)人HepG2細(xì)胞中Rb和MAD2表達(dá)的影響[J];中國(guó)病理生理雜志;2014年05期

4 丁浩;張吉翔;;人著色性干皮病基因D對(duì)HepG2細(xì)胞Mdm2和Mdm4表達(dá)的影響[J];中國(guó)病理生理雜志;2014年01期

5 Yong-Jun Fan;Wei-Xing Zong;;The cellular decision between apoptosis and autophagy[J];Chinese Journal of Cancer;2013年03期

6 鄒穎穎;張吉翔;;人剪切修復(fù)基因XPD對(duì)肝癌細(xì)胞生長(zhǎng)及ERG基因的影響[J];天津醫(yī)藥;2013年01期

7 王娟;杜芳騰;杜瑤;張吉翔;;人剪切修復(fù)基因XPD對(duì)人肝癌SMMC-7721細(xì)胞中Ets-1和Cdk6基因的調(diào)控作用[J];天津醫(yī)藥;2012年09期

8 何月;羅文;張吉翔;;XPD和PPARγ對(duì)HepG2增殖及ERG表達(dá)的影響[J];醫(yī)學(xué)分子生物學(xué)雜志;2012年04期

9 馬果果;張吉翔;;人剪切修復(fù)基因XPD對(duì)p52和p21基因的影響[J];天津醫(yī)藥;2011年06期

10 湯蕾;張吉翔;熊瑛;;人著色性干皮病D組基因的克隆及其真核表達(dá)[J];生物醫(yī)學(xué)工程學(xué)雜志;2008年03期

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