【摘要】：在人乳腺癌細(xì)胞系BT549中穩(wěn)定干擾整合素β1(integrinβ1,ITGB1),研究其對乳腺癌細(xì)胞遷移和侵襲的影響。構(gòu)建靶向干擾ITGB1基因的sh RNA慢病毒,感染BT549細(xì)胞,通過篩選成功獲得穩(wěn)定干擾ITGB1蛋白的BT-549細(xì)胞系,實(shí)驗(yàn)設(shè)空白對照組(Blank)、病毒感染陰性對照組(shNC)、ITGB1-shRNA慢病毒感染組(shITGB1);Real-time PCR和Western blotting法檢測穩(wěn)定干擾后ITGB1 mRNA和蛋白的表達(dá)情況;Western blotting法檢測穩(wěn)定干擾ITGB1后上皮間質(zhì)轉(zhuǎn)化(epithelial-mesenchymal transition,EMT)相關(guān)標(biāo)志蛋白(E-cadherin,Vimentin,N-cadherin,Fibronectin)的表達(dá)情況;利用劃痕試實(shí)驗(yàn)和Transwell侵襲試實(shí)驗(yàn)分別檢測穩(wěn)定干擾ITGB1后細(xì)胞的遷移及侵襲能力。Real-time PCR和Western blotting結(jié)果顯示,shITGB1組細(xì)胞ITGB1 mRNA和蛋白的表達(dá)水平顯著低于Blank組和shNC組細(xì)胞(p0.01,p0.01)。Western blotting結(jié)果顯示,穩(wěn)定干擾ITGB1可部分逆轉(zhuǎn)乳腺癌細(xì)胞BT549的EMT化;穩(wěn)定干擾ITGB1后,上皮標(biāo)志蛋白E-cadherin表達(dá)顯著上調(diào)(p0.01),間質(zhì)標(biāo)志蛋白Vimentin(p0.05)、N-cadherin(p0.01)和Fibronectin(p0.01)的表達(dá)顯著降低。劃痕和Transwell侵襲試驗(yàn)實(shí)驗(yàn)結(jié)果顯示,穩(wěn)定干擾ITGB1后可顯著降低乳腺癌細(xì)胞的遷移及侵襲能力(p0.05,p0.05)。在乳腺癌BT549細(xì)胞中穩(wěn)定干擾ITGB1后,通過逆轉(zhuǎn)EMT化抑制細(xì)胞遷移和侵襲。
[Abstract]:To study the effects of integrin 尾 1-ITGB1 on the migration and invasion of human breast cancer cell line BT549. Sh RNA lentivirus targeting ITGB1 gene was constructed and infected with BT549 cells. BT-549 cell lines with stable interfering ITGB1 protein were obtained by screening successfully. Real-time PCR and Western blotting methods were used to detect the expression of ITGB1 mRNA and protein after stable interference in the control group with negative (Blank), virus infection. Western blotting assay was used to detect epithelial-mesenchymal transition EMT related to stable interfering ITGB1. The expression of E-cadherin in N-cadherin fibronectin; The results of scratch test and Transwell invasion test showed that the expression of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01, p0.01). Western blotting showed that the expression level of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01). The results of real-time PCR and Western blotting showed that the expression of ITGB1 mRNA and protein was significantly lower than that of Blank group and shNC group (p0.01). Stable interference with ITGB1 could partially reverse the EMT expression of BT549 in breast cancer cells, and the expression of E-cadherin, Vimentin (p0.05), N-cadherin (p0.01) and Fibronectin (p0.01) were significantly up-regulated (p0.01), and the expression of Vimentin (p0.01) and Fibronectin (p0.01) were significantly decreased. The results of scratch and Transwell invasion tests showed that stable interference with ITGB1 could significantly reduce the migration and invasion ability of breast cancer cells (p0.05). Stable interference with ITGB1 in breast cancer BT549 cells inhibited migration and invasion by reversing EMT.
【作者單位】： 重慶醫(yī)科大學(xué)檢驗(yàn)醫(yī)學(xué)院臨床檢驗(yàn)診斷學(xué)教育部重點(diǎn)實(shí)驗(yàn)室;成都市第三人民醫(yī)院檢驗(yàn)科;
【基金】：國家自然科學(xué)基金(31171336)項(xiàng)目資助
【分類號(hào)】：R737.9

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