本文選題：膠質(zhì)瘤 + LRIG1��； 參考：《武漢大學(xué)》2016年博士論文

【摘要】：目的：探討LRIG1對(duì)人腦膠質(zhì)瘤細(xì)胞侵襲及轉(zhuǎn)移能力的影響及其可能的分子機(jī)制。探討SNAI2|—E-cadherin軸在這一過(guò)程中所發(fā)揮的作用。方法：使用質(zhì)粒轉(zhuǎn)染LRIG1基因的方法使人腦膠質(zhì)瘤細(xì)胞系U251細(xì)胞中高表達(dá)LRIG1,采用G418篩選出陽(yáng)性單克隆細(xì)胞株并進(jìn)行擴(kuò)增,RT-PCR及Western Blot驗(yàn)證LRIG1在mRNA及蛋白水平高表達(dá)。應(yīng)用siRNA使人腦膠質(zhì)瘤細(xì)胞系U251細(xì)胞中低表達(dá)LRIG1,RT-PCR及Western Blot驗(yàn)證LRIG1在mRNA及蛋白水平低表達(dá)。Transwell侵襲實(shí)驗(yàn)檢測(cè)細(xì)胞侵襲能力的變化,Transwell轉(zhuǎn)移實(shí)驗(yàn)及劃痕愈合實(shí)驗(yàn)檢測(cè)細(xì)胞轉(zhuǎn)移能力的變化。RT-PCR及Western Blot檢測(cè)LRIG1表達(dá)水平改變時(shí)EMT相關(guān)標(biāo)記分子SNAI2及其下游分子E-cadherin的變化。收集人腦膠質(zhì)母細(xì)胞瘤及人正常腦組織標(biāo)本,采用Western Blot檢測(cè)各例標(biāo)本中LRIG1與SNAI2和E-cadherin的蛋白表達(dá)。最后使用SNAI2的小分子抑制劑PCPA處理LRIG1低表達(dá)的U251細(xì)胞,觀察SNAI2功能被抑制后低表達(dá)LRIG1對(duì)細(xì)胞侵襲及轉(zhuǎn)移能力的影響及SNAI2—E-cadherin軸的變化。結(jié)果：運(yùn)用質(zhì)粒轉(zhuǎn)染的方法成功構(gòu)建高表達(dá)LRIG1的人腦膠質(zhì)瘤U251細(xì)胞株。LRIG1高表達(dá)可導(dǎo)致U251細(xì)胞侵襲及轉(zhuǎn)移能力明顯下降。同時(shí)LRIG1高表達(dá)會(huì)下調(diào)細(xì)胞中SNAI2的表達(dá),導(dǎo)致細(xì)胞中E-cadherin表達(dá)量明顯上升。運(yùn)用特異性沉默LRIG1的siRNA成功構(gòu)建低表達(dá)LRIG1的人腦膠質(zhì)瘤U251細(xì)胞株。LRIG1低表達(dá)可導(dǎo)致U251細(xì)胞侵襲及轉(zhuǎn)移能力明顯上升。同時(shí)LRIG1低表達(dá)會(huì)上調(diào)細(xì)胞中SNAI2的表達(dá),導(dǎo)致細(xì)胞中E-cadherin表達(dá)量明顯下降。與正常人腦組織相比,人腦膠質(zhì)母細(xì)胞瘤組織中LRIG1蛋白水平較低,SNAI2蛋白水平明顯上調(diào)而E-cadherin蛋白水平明顯下調(diào)。進(jìn)一步實(shí)驗(yàn)表明,使用PCPA處理LRIG1低表達(dá)的U251細(xì)胞阻斷SNAI2的功能,可逆轉(zhuǎn)LRIG1低表達(dá)對(duì)U251細(xì)胞侵襲和轉(zhuǎn)移能力的促進(jìn)作用及對(duì)E-caherin表達(dá)的調(diào)節(jié)作用。結(jié)論：LRIG1可以抑制人腦膠質(zhì)瘤U251細(xì)胞侵襲及轉(zhuǎn)移,這一作用與LRIG1抑制SNAI2—E-cadherin軸有關(guān)。人腦膠質(zhì)母細(xì)胞瘤LRIG1蛋白水平較低,SNAI2蛋白水平較高,E-cadherin蛋白水平較低。人腦膠質(zhì)母細(xì)胞瘤中LRIG1可通過(guò)SNAI2—E-cadherin軸調(diào)節(jié)細(xì)胞侵襲與轉(zhuǎn)移能力,但其詳細(xì)的分子機(jī)制尚有待進(jìn)一步研究。
[Abstract]:Aim: to investigate the effect of LRIG1 on invasion and metastasis of human glioma cells and its possible molecular mechanism.The role of SNAI2-E-cadherin axis in this process is discussed.Methods: the expression of LRIG1 in human glioma cell line U251 was induced by plasmid transfection of LRIG1 gene. The positive monoclonal cell line was screened by G418, and the overexpression of LRIG1 in mRNA and protein levels was verified by RT-PCR and Western Blot.Using siRNA to induce low expression of LRIG1T in human glioma cell line U251 by RT-PCR and Western Blot to verify the change of cell invasion ability of LRIG1 in mRNA and protein low expression .Transwell invasion assay; Transwell metastasis test; scratch healing assay to detect cell metastasisAbility change. RT-PCR and Western Blot were used to detect the changes of EMT related marker SNAI2 and its downstream E-cadherin.Western Blot was used to detect the expression of LRIG1, SNAI2 and E-cadherin in human glioblastoma and normal brain tissue.Finally, U251 cells with low expression of LRIG1 were treated with PCPA, a small molecular inhibitor of SNAI2. The effect of low expression of LRIG1 on the invasion and metastasis of U251 cells and the changes of SNAI2-E-cadherin axis were observed after SNAI2 function was inhibited.Results: the overexpression of human glioma cell line. LRIG1 was successfully constructed by plasmid transfection. The ability of invasion and metastasis of U251 cells decreased significantly.At the same time, the high expression of LRIG1 can down-regulate the expression of SNAI2, leading to a significant increase in the expression of E-cadherin.The low expression of human glioma cell line. LRIG1, which was successfully constructed by using siRNA with specific silencing of LRIG1, could significantly increase the ability of invasion and metastasis of U251 cells.At the same time, low expression of LRIG1 upregulated the expression of SNAI2, which resulted in the decrease of E-cadherin expression.Compared with normal human brain tissue, the level of LRIG1 protein in glioblastoma tissue was lower than that in normal human brain tissue. The level of SNAI2 protein was up-regulated and the level of E-cadherin protein was down-regulated.Further experiments showed that PCPA treatment of U251 cells with low expression of LRIG1 could reverse the effect of LRIG1 low expression on the invasion and metastasis of U251 cells and the regulation of E-caherin expression by blocking the function of SNAI2.Conclusion: LRIG1 can inhibit the invasion and metastasis of human glioma U251 cells, which is related to the inhibition of SNAI2-E-cadherin axis by LRIG1.The level of LRIG1 protein in human glioblastoma was lower and SNAI2 protein level was higher than that of E-cadherin protein.LRIG1 can regulate the invasion and metastasis of human glioblastoma through the SNAI2-E-cadherin axis, but the detailed molecular mechanism remains to be further studied.
【學(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2016
【分類號(hào)】：R739.41

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 夏紅強(qiáng);E-cadherin在腫瘤組織的檢測(cè)及意義[J];臨床腫瘤學(xué)雜志;2003年06期

2 ;Expression of TGF-β1,Snail,E-cadherin and N-cadherin in Gastric Cancer and Its Significance[J];Chinese Journal of Clinical Oncology;2007年06期

3 熊小蔚;崔明;;T-cadherin與腫瘤[J];醫(yī)學(xué)綜述;2010年08期

4 ;E－Cadherin在膀胱乳頭狀移行細(xì)胞癌的表達(dá)[J];國(guó)外醫(yī)學(xué)(生理、病理科學(xué)與臨床分冊(cè));1996年02期

5 張文軍,張志謙,林仲翔,王耐勤,胡穎;Correlation between expression of cadherin and gap junctional communication in human lung carcinoma cells[J];Science in China(Series C:Life Sciences);1998年04期

6 ;The expression of N-cadherin/catenins/actin complex in human lung normal and carcinoma cells[J];Chinese Science Bulletin;1998年09期

7 ;Quantitative analysis of E-cadherin expression and clinicopathologic evaluation in gastric cancer[J];Chinese Medical Journal;1998年01期

8 Ge-Liang Liu;Han-Jie Yang;Tian Liu;Yun-Zhao Lin;;Expression and significance of E-cadherin,N-cadherin,transforming growth factor-β1 and Twist in prostate cancer[J];Asian Pacific Journal of Tropical Medicine;2014年01期

9 王曦,吳人亮,郝天玲,陳芳;Effects of Cigarette Smoke Extract on E-cadherin Expression in Cultured Airway Epithelial Cells[J];Journal of Tongji Medical University;2000年01期

10 陳曉峰,丁嘉安,高文,易祥華,王海峰,李化;E-cadherin表達(dá)與非小細(xì)胞肺癌淋巴結(jié)轉(zhuǎn)移及預(yù)后的相關(guān)性研究[J];中國(guó)肺癌雜志;2002年04期

相關(guān)會(huì)議論文 前10條

1 ;Core Fucosylation Regulates E-cadherin Binding Affinity in Human Lung Cancer Cells[A];2008年全國(guó)生物化學(xué)與分子生物學(xué)學(xué)術(shù)大會(huì)論文摘要[C];2008年

2 ;Epigenetic regulation of E-cadherin in HBV associate human Hepatocellular carcinoma[A];江蘇省遺傳學(xué)會(huì)第八屆會(huì)員代表大會(huì)暨學(xué)術(shù)研討會(huì)論文集[C];2010年

3 王冰晶;張志謙;;癌細(xì)胞cadherin亞型轉(zhuǎn)變及其分子機(jī)制[A];中國(guó)細(xì)胞生物學(xué)學(xué)會(huì)2005年學(xué)術(shù)大會(huì)、青年學(xué)術(shù)研討會(huì)論文摘要集[C];2005年

4 ;Suppressed survival and proliferation of ovarian cancer cells by repressed MEK-ERK activation via knocking-down of E-cadherin[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

5 ;N-cadherin:a potential receptor of GDNF[A];Proceedings of the 8th Biennial Conference of the Chinese Society for Neuroscience[C];2009年

6 Zhiyang Xu;Dechang Li;Baohua Ji;;Quantification of the stiffness and strength of cadherin ectodomain binding with different ions[A];北京力學(xué)會(huì)第20屆學(xué)術(shù)年會(huì)論文集[C];2014年

7 賈文亮;張慶瑜;;關(guān)于E-cadherin在胃癌和肺癌中表達(dá)情況的研究[A];天津市生物醫(yī)學(xué)工程學(xué)會(huì)第三十三屆學(xué)術(shù)年會(huì)論文集[C];2013年

8 烏新春;倪志超;段昕所;宋有鑫;;綠色熒光蛋白標(biāo)記的T-cadherin基因穩(wěn)定轉(zhuǎn)染黑素瘤細(xì)胞株的建立[A];第五屆全國(guó)中醫(yī)藥免疫學(xué)術(shù)研討會(huì)——暨環(huán)境·免疫與腫瘤防治綜合交叉會(huì)議論文匯編[C];2009年

9 ;Manipulation of N-cadherin level and function through different methods results in distinct functional changes in hippocampal neurons[A];Proceedings of the 7th Biennial Meeting and the 5th Congress of the Chinese Society for Neuroscience[C];2007年

10 張憲真;陳隨芹;耿秀東;;N-Cadherin與E-Cadherin在非小細(xì)胞肺癌中的表達(dá)及相關(guān)性研究[A];中國(guó)腫瘤內(nèi)科進(jìn)展 中國(guó)腫瘤醫(yī)師教育（2014）[C];2014年

相關(guān)博士學(xué)位論文 前10條

1 楊俊俊;Rab5 GTPase介導(dǎo)的VE-cadherin內(nèi)吞調(diào)控肺微血管內(nèi)皮細(xì)胞屏障功能的機(jī)制[D];第三軍醫(yī)大學(xué);2015年

2 張紅燕;STAT3/ZEB1/E-cadherin信號(hào)軸在食管鱗癌上皮間質(zhì)轉(zhuǎn)化中的作用及機(jī)制研究[D];鄭州大學(xué);2016年

3 陶祥;SNAI2—E-cadherin軸在LRIG1調(diào)節(jié)人腦膠質(zhì)瘤細(xì)胞侵襲和轉(zhuǎn)移中的作用研究[D];武漢大學(xué);2016年

4 黎景佳;E-cadherin在喉鱗狀細(xì)胞癌中的表達(dá)及其與腫瘤侵襲轉(zhuǎn)移的關(guān)系[D];中南大學(xué);2010年

5 吳揚(yáng);VE-cadherin重組腺病毒偶聯(lián)氧化型甘露聚糖抗腫瘤實(shí)驗(yàn)研究[D];四川大學(xué);2005年

6 胡軍;E-cadherin與卵巢癌轉(zhuǎn)移的相關(guān)性及機(jī)制研究[D];大連醫(yī)科大學(xué);2007年

7 來(lái)明名;肌球蛋白X與N-cadherin相互作用調(diào)節(jié)皮層神經(jīng)元輻射遷移[D];東北師范大學(xué);2013年

8 任劍珍;結(jié)腸癌T-cadherin基因的表達(dá)及甲基化影響的探討[D];中南大學(xué);2011年

9 張自森;E-cadherin在胃癌中的表達(dá)與化療療效的關(guān)系及對(duì)胃癌細(xì)胞化療敏感性的影響[D];鄭州大學(xué);2013年

10 余瓊芳;Li-cadherin與結(jié)腸癌LoVo細(xì)胞侵襲轉(zhuǎn)移相關(guān)性實(shí)驗(yàn)研究[D];武漢大學(xué);2009年

相關(guān)碩士學(xué)位論文 前10條

1 衷燦梅;12-脂氧合酶通過(guò)上皮間質(zhì)轉(zhuǎn)化促進(jìn)胃癌侵襲轉(zhuǎn)移的機(jī)制研究[D];福建醫(yī)科大學(xué);2015年

2 高玉梅;探索N-cadherin在卵巢癌發(fā)生和進(jìn)展中的作用及意義[D];石河子大學(xué);2015年

3 林路;E-cadherin,β-catenin,N-cadherin在漿液性卵巢癌中的表達(dá)及意義[D];安徽醫(yī)科大學(xué);2015年

4 辛明;Galectin-3通過(guò)EGFR介導(dǎo)調(diào)控腫瘤細(xì)胞E-cadherin表達(dá)的機(jī)制研究[D];山東大學(xué);2015年

5 孟梅;VE-cadherin對(duì)血管穩(wěn)態(tài)的作用及調(diào)控機(jī)制[D];蘇州大學(xué);2015年

6 汪威;丙泊酚對(duì)荷瘤大鼠MADB106腫瘤細(xì)胞肺轉(zhuǎn)移及E-cadherin和β-catenin的影響[D];南方醫(yī)科大學(xué);2015年

7 孫琳;ALCAM、E-cadherin和β-catenin在甲狀腺乳頭狀癌中的表達(dá)及意義[D];河北醫(yī)科大學(xué);2014年

8 于海峰;FOXQ1和E-cadherin在食管鱗狀細(xì)胞癌中的表達(dá)及臨床意義[D];天津醫(yī)科大學(xué);2015年

9 何宏月;EMT相關(guān)蛋白E-cadherin、N-cadherin、Snail和Slug在子宮內(nèi)膜異位癥中的表達(dá)及意義[D];天津醫(yī)科大學(xué);2015年

10 劉超;EMX2和E-cadherin在食管鱗癌中的表達(dá)及臨床病理意義[D];天津醫(yī)科大學(xué);2015年

，

本文編號(hào)：1762409