本文選題：siRNA + RhoGDI�。� 參考：《中國(guó)免疫學(xué)雜志》2017年04期

【摘要】：目的:應(yīng)用小分子干擾RNA(siRNA)沉默RhoGDI2基因的表達(dá),初步探討其對(duì)結(jié)腸癌細(xì)胞增殖、遷移能力的影響及可能的機(jī)制。方法:分別運(yùn)用Western blot和RT-q PCR檢測(cè)結(jié)腸癌細(xì)胞株RKO、HT29、SW620、SW480、HCT116基因RhoGDI2的表達(dá)情況。設(shè)計(jì)并合成RhoGDI2 siRNA干擾序列,按照LipofectamineTM2000轉(zhuǎn)染方法將siRNA干擾序列轉(zhuǎn)染到目的細(xì)胞,設(shè)置實(shí)驗(yàn)干擾組、空白對(duì)照和陰性對(duì)照組;CCK-8實(shí)驗(yàn)檢測(cè)細(xì)胞增殖能力,細(xì)胞劃痕試驗(yàn)和Transwell實(shí)驗(yàn)檢測(cè)細(xì)胞遷移、侵襲能力。結(jié)果:人結(jié)腸癌細(xì)胞株RhoGDI2表達(dá)量由高到低依次是RKO、HT29、SW620、SW480、HCT116;RKO細(xì)胞siRNA干擾后RhoGDI2表達(dá)抑制率大于70%:實(shí)驗(yàn)干擾組、陰性對(duì)照組、空白對(duì)照組細(xì)胞增殖率分別是(0.683±0.013)、(0.866±0.088)、(0.905±0.008),P0.05;實(shí)驗(yàn)干擾組細(xì)胞遷移、侵襲速率較對(duì)照組均減慢。沉默RhoGDI2基因的表達(dá),實(shí)驗(yàn)組細(xì)胞E-Cadherin較對(duì)照組表達(dá)量高,Vimentin蛋白表達(dá)下降。結(jié)論:結(jié)腸癌RhoGDI2基因沉默可能通過(guò)抑制EMT進(jìn)程阻止腫瘤惡性生物學(xué)行為。
[Abstract]:Aim: to silence the expression of RhoGDI2 gene by small interfering RNAs (siRNAs), and to explore the possible mechanism of its effect on the proliferation and migration of colon cancer cells. Methods: Western blot and RT-q PCR were used to detect the RhoGDI2 expression of HCT116 gene. The RhoGDI2 siRNA interference sequence was designed and synthesized, and the siRNA interference sequence was transfected into the target cell according to the LipofectamineTM2000 transfection method. The experimental interference group, the blank control group and the negative control group were used to detect the proliferation ability of the cells. Cell scratch test and Transwell assay were used to detect cell migration and invasion. Results: the order of RhoGDI2 expression in human colon cancer cell line was RKOHT29, SW620, SW480, HCT116, RKO cells, the inhibition rate of RhoGDI2 expression was higher than 70: the proliferative rate of experimental interference group, negative control group and blank control group was 0. 683 鹵0. 083 鹵0. 0888 鹵0. 905 鹵0. 008P0.05. the cell migration of experimental interference group, negative control group and blank control group were 0. 683 鹵0. 088 鹵0. 905 鹵0. 008P0. 05 respectively. The invasion rate was slower than that of the control group. Silencing the expression of RhoGDI2 gene, the expression of Vimentin protein in the experimental group was significantly lower than that in the control group. Conclusion: RhoGDI2 gene silencing in colon cancer may prevent malignant biological behavior by inhibiting EMT progression.
【作者單位】： 吉林大學(xué)第二醫(yī)院肝膽胰內(nèi)科;第二軍醫(yī)大學(xué)長(zhǎng)海醫(yī)院檢驗(yàn)科;上海交通大學(xué)醫(yī)學(xué)院附屬同仁醫(yī)院腫瘤科;
【基金】：吉林省科技廳科研基金課題(2013c026-1)
【分類(lèi)號(hào)】：R735.35

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