本文關(guān)鍵詞：LIM家族蛋白Ajuba對(duì)腫瘤細(xì)胞的生物學(xué)功能與分子機(jī)制研究　出處：《上海交通大學(xué)》2015年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： Ajuba Twist 細(xì)胞遷移 結(jié)直腸癌 Ajuba 雌激素受體α 乳腺癌 細(xì)胞生長(zhǎng)

【摘要】：目的探索結(jié)直腸癌細(xì)胞中Ajuba與Twist之間的相互作用,及該作用對(duì)于結(jié)直腸癌遷移所產(chǎn)生的影響。方法運(yùn)用免疫共沉淀方法證明外源性Ajuba與Twist蛋白質(zhì)之間存在相互作用;熒光素酶活性檢測(cè)Ajuba對(duì)于Twist下游靶基因N-cadherin啟動(dòng)子的轉(zhuǎn)錄活性;構(gòu)建結(jié)直腸癌SW1116-sh Ajuba/OEAjuba穩(wěn)轉(zhuǎn)細(xì)胞,并進(jìn)行Transwell侵襲實(shí)驗(yàn)觀察細(xì)胞遷移能力變化。結(jié)果Ajuba與Twsit蛋白質(zhì)之間存在相互作用,并且Ajuba能通過Twist促進(jìn)N-cadherin的表達(dá)調(diào)控。在結(jié)直腸癌細(xì)胞SW1116-sh Ajuba穩(wěn)轉(zhuǎn)細(xì)胞中可發(fā)現(xiàn),AJUBA基因表達(dá)水平降低,蛋白表達(dá)下降,能抑制細(xì)胞遷移能力。結(jié)論Ajuba是轉(zhuǎn)錄因子Twist的共活化因子(coactivator),能夠促進(jìn)N-cadherin表達(dá),增強(qiáng)結(jié)直腸癌細(xì)胞的遷移能力。目的探索Ajuba在雌激素受體α(Estrogen receptorα,ERα)陽(yáng)性的乳腺癌細(xì)胞中的生物學(xué)作用,依據(jù)已知的Ajuba與ERα存在相互作用尋找涉及的分子機(jī)制。方法運(yùn)用免疫共沉淀方法證明外源性Ajuba與ERα蛋白質(zhì)之間存在相互作用;構(gòu)建乳腺癌T47D-sh Ajuba、T47D-oe Ajuba穩(wěn)轉(zhuǎn)細(xì)胞,并進(jìn)行CCK8實(shí)驗(yàn)觀察細(xì)胞生長(zhǎng)能力變化;建立T47D-sh ERα穩(wěn)轉(zhuǎn)細(xì)胞,比較生長(zhǎng)趨勢(shì)改變;熒光素酶活性檢測(cè)Ajuba-ERα對(duì)于下游靶基因CDK6啟動(dòng)子的轉(zhuǎn)錄活性調(diào)控。結(jié)果Ajuba與ERα之間存在相互作用,并且Ajuba能通過ERα促進(jìn)CDK6的表達(dá)調(diào)控。在乳腺癌細(xì)胞T47D-sh Ajuba穩(wěn)轉(zhuǎn)細(xì)胞中可發(fā)現(xiàn),AJUBA基因表達(dá)水平降低,蛋白表達(dá)下降,細(xì)胞生長(zhǎng)緩慢;細(xì)胞中過表達(dá)Ajuba,則可上調(diào)CDK6的表達(dá)水平。結(jié)論Ajuba是ERα的共活化因子(coactivator),能夠促進(jìn)CDK6表達(dá),參與調(diào)控乳腺癌細(xì)胞T47D的生長(zhǎng)。
[Abstract]:Objective to explore the interaction between Ajuba and Twist in colorectal cancer cells. Methods Immunoprecipitation method was used to prove the interaction between exogenous Ajuba and Twist protein. Luciferase activity was used to detect the transcriptional activity of Ajuba on N-cadherin promoter of Twist downstream gene. Construction of SW1116-sh Ajuba/OEAjuba stable transformed cells from colorectal cancer. The ability of cell migration was observed by Transwell invasion assay. Results there was interaction between Ajuba and Twsit protein. Ajuba can promote the regulation of N-cadherin expression through Twist. It can be found in SW1116-sh Ajuba stable transformed cells. The decrease of AJUBA gene expression and the decrease of protein expression can inhibit the ability of cell migration. Conclusion Ajuba is the coactivator of transcription factor Twist. It can promote N-cadherin expression and enhance the migration ability of colorectal cancer cells. Objective to explore the role of Ajuba in estrogen receptor 偽 strogen receptor 偽. The biological role of ER 偽 in breast cancer cells. Methods the interaction between exogenous Ajuba and ER 偽 protein was demonstrated by immunoprecipitation method. Breast cancer T47D-sh Ajubahe T47D-OE Ajuba was constructed, and the growth ability was observed by CCK8 assay. T47D-sh ER 偽 stable cells were established to compare the growth trend. Luciferase activity was used to detect the transcriptional regulation of Ajuba-ER 偽 on the downstream target gene CDK6 promoter. Results there was an interaction between Ajuba and ER 偽. Moreover, Ajuba can promote the regulation of CDK6 expression through ER 偽. The decrease of AJUBA gene expression can be found in T47D-sh Ajuba stable transfer cells of breast cancer cells. Protein expression decreased and cell growth slowed down. Overexpression of Ajuba can up-regulate the expression of CDK6. Conclusion Ajuba is a coactivator of ER 偽 and can promote the expression of CDK6. Participate in regulating the growth of breast cancer cell T47D.
【學(xué)位授予單位】：上海交通大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R735.34

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