【摘要】：目的:探討細(xì)胞周期蛋白D1(cyclin D1)的編碼基因CCND1 miR-340介導(dǎo)的逆轉(zhuǎn)結(jié)直腸癌細(xì)胞對5-氟尿嘧啶(5-Fu)耐藥的機(jī)制。方法:采用瞬時轉(zhuǎn)染技術(shù)將結(jié)直腸癌細(xì)胞HCT116、SW480株分別轉(zhuǎn)染si-CCND1和miR340-mimic。應(yīng)用MTT法檢測轉(zhuǎn)染后的結(jié)直腸癌細(xì)胞對5-Fu敏感性的變化,應(yīng)用雙熒光素酶試驗驗證CCND1對miR340參與的影響結(jié)直腸癌細(xì)胞對5-Fu敏感性的影響。結(jié)果:瞬時轉(zhuǎn)染si CCND1和過表達(dá)miR-340后,結(jié)直腸癌HCT116和SW480細(xì)胞的IC50值均顯著低于對照組(10,10 vs 20μmol/L和20,20 vs 40μmol/L,均P0.05)。共轉(zhuǎn)染CCND1 3'UTR野生質(zhì)粒和miR-340 inhibitor的結(jié)直腸癌HCT116和SW48細(xì)胞熒光素酶的活性顯著高于共轉(zhuǎn)染空載體和mimic細(xì)胞(P0.01)。結(jié)論:CCND1作為不良因子通過抑制miR340的表達(dá)進(jìn)而發(fā)揮增加結(jié)直腸癌細(xì)胞對5-Fu耐藥的作用。
[Abstract]:Aim: to investigate the mechanism of reversal of 5-fluorouracil (5-Fu) resistance in colorectal cancer cells mediated by cyclin D1 (cyclin D1) encoding gene CCND1 miR-340. Methods: HCT116,SW480 cells were transfected into si-CCND1 and miR340-mimic. by transient transfection technique. MTT assay was used to detect the sensitivity of transfected colorectal cancer cells to 5-Fu, and double luciferase assay was used to verify the effect of CCND1 on the 5-Fu sensitivity of colorectal cancer cells involved in miR340. Results: after transient transfection of si CCND1 and overexpression of miR-340, the IC50 values of HCT116 and SW480 cells in colorectal cancer were significantly lower than those in control group (10 10 vs 20 渭 mol/L and 20 20 vs 40 渭 mol/L, P0.05). The luciferase activity of HCT116 and SW48 cells cotransfected with CCND1 3'UTR wild plasmid and miR-340 inhibitor was significantly higher than that of empty vector and mimic cells (P0.01). Conclusion as an adverse factor, the expression of miR340 is inhibited and the resistance to 5-Fu is increased in colorectal cancer cells.
【作者單位】： 廣州中醫(yī)藥大學(xué)附屬海南省中醫(yī)院腫瘤科;南方醫(yī)科大學(xué)中西醫(yī)結(jié)合醫(yī)院腫瘤中心;
【基金】：廣東省自然科學(xué)基金資助項目(No.2015A030311005)~~
【分類號】：R735.34

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