MicroRNA-155在肝細胞癌對索拉非尼抗藥中的作用研究
發(fā)布時間:2018-06-22 19:59
本文選題:微小RNA + 肝細胞癌。 參考:《中國藥理學通報》2017年05期
【摘要】:目的探討微小RNA155(microRNA-155,miR-155)在肝細胞癌(hepatocellular carcinoma,HCC)對索拉非尼(sorafenib)抗藥中的作用。方法將miR-155抑制慢病毒(miR-155 inhibitor)轉(zhuǎn)染miR-155表達相對較高的SMMC-7721細胞,而miR-155過表達慢病毒(miR-155)轉(zhuǎn)染miR-155表達相對較低的Hep G2細胞;用熒光定量PCR(qPCR)檢測經(jīng)慢病毒轉(zhuǎn)染的SMMC-7721細胞及HepG2細胞miR-155表達量,以驗證慢病毒轉(zhuǎn)染效果;通過CCK-8法及流式細胞術(shù)檢測各組細胞經(jīng)索拉非尼作用后的存活率及凋亡情況;用Western blot檢測凋亡相關(guān)蛋白活性caspase-3表達量,從而進一步探究各組細胞凋亡情況。結(jié)果與對照組相比,轉(zhuǎn)染miR-155抑制慢病毒的SMMC-7721細胞表達miR-155明顯下調(diào)(P0.01),經(jīng)索拉非尼處理后其存活率明顯降低(P0.05),而索拉非尼誘導其凋亡明顯增加(P0.01),其活性caspase-3表達量明顯上調(diào)(P0.01);miR-155過表達慢病毒轉(zhuǎn)染HepG2細胞后,則相反。結(jié)論 miR-155參與肝細胞癌對索拉非尼的抗藥,有希望成為一個肝癌治療的新靶標。
[Abstract]:Objective to investigate the role of microRNA-155 miR-155 in the resistance of hepatocellular carcinoma to Solafenil (sorafenib). Methods miR-155 inhibited lentivirus (miR-155 inhibitor) was transfected into SMMC-7721 cells with relatively high miR-155 expression, while miR-155 overexpression lentivirus (miR-155) was transfected into Hep G2 cells with relatively low miR-155 expression. The expression of miR-155 in SMMC-7721 cells and HepG2 cells was detected by fluorescence quantitative polymerase chain reaction (qPCR). In order to verify the effect of lentivirus transfection, CCK-8 and flow cytometry were used to detect the survival rate and apoptosis of the cells treated with Solafenil, and Western blot was used to detect the expression of apoptosis-related protein (caspase-3). So as to further explore the apoptosis of each group. Results compared with the control group, The expression of miR-155 was down-regulated (P0.01) in SMMC-7721 cells transfected with miR-155. The survival rate of SMMC-7721 cells treated with sorafenil was significantly decreased (P0.05), while the apoptosis induced by sorafenil was significantly increased (P0.01). The expression of active caspase-3 was up-regulated (P0.01) and the overexpression of miR-155 was up-regulated (P0.01) in HepG2 cells. The opposite is true. Conclusion miR-155 may be a new target for the treatment of hepatocellular carcinoma.
【作者單位】: 重慶醫(yī)科大學附屬第一醫(yī)院肝膽外科;
【基金】:國家自然科學基金資助項目(No 81470898)
【分類號】:R735.7
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