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1、·828·中華實驗外科雜志2014年4月第31卷第4期ChinJExpSurg，April2014，Vo1．31，No．4·實驗研究·腫瘤干細(xì)胞在人骨肉瘤細(xì)胞株MG63耐JJb~,Cn效應(yīng)中的作用張明生劉萍萍朱宇張仲寧陳清漢韓娜【摘要】目的觀察腫瘤干細(xì)胞在人骨肉瘤細(xì)胞株對順鉑(CDDP)產(chǎn)生獲得性耐藥中的作用。方法采用大劑量沖擊結(jié)合逐步增加劑量(1、3、5mg／L依次遞增)的方法誘導(dǎo)產(chǎn)生人骨肉瘤耐CDDP細(xì)胞株，0．20mg／L的As，O逆轉(zhuǎn)耐藥株的耐藥性，噻唑藍(lán)(MTT)法和實時定量聚合酶鏈反應(yīng)(Re
2、al—timePCR)分別檢測細(xì)胞株對CDDP的敏感性和let一7、微小RNA(miR)一200c以及性別決定區(qū)Y框蛋白一2(SOX一2)的表達(dá)。結(jié)果耐藥株的耐藥性顯著高于親本株(P<0．01)。As，O降低了耐藥株對CDDP的耐藥性，逆轉(zhuǎn)倍數(shù)為3．39，相對逆轉(zhuǎn)效率為76．6％。相對于親本株，耐藥株細(xì)胞let-7和miR-200c的表達(dá)顯著降低(P<0．05)，SOX一2的表達(dá)顯著升高(P<0．01)。AsO處理后，耐藥株細(xì)胞let-7和miR一200c的表達(dá)顯著增加(P<0．05)，SOX一2的表達(dá)
3、顯著下降(P<0．05)。結(jié)論CSC有可能是人骨肉瘤細(xì)胞株MG63對CDDP產(chǎn)生耐藥的機制之一。【關(guān)鍵詞】骨肉瘤；耐藥；順鉑；腫瘤干細(xì)胞TheroleofcancerstemcellsintheacquiredresistancetocisplatinofhumanosteosarcomacelllineMG63ZhangMingsheng，LiuPingping，ZhuYu，ZhangZhongning，ChenQinghan，HanNa．DepartmentofOrthopaedics，theSec
4、ondAffiliatedHospitalofZhengzhouUniversity，Zhengzhou450014，ChinaCorrespondingauthor：HanNn，Email：592501251@qq．corn【Abstract】0bjectiveToinvestigatetheroleofcancerstemceils(CSC)intheacquiredresist—ancetocisplatinfCDDP)ofhumanosteosareomaeelllineMG63．Methods
5、CDDP—resistanthumanos—teosarcomacelllineM(3fMG63／CDDP)wasestablishedvitrobyexposuretohighandgraduallyin—creaseddoseofcisplatin(13，5Ing／L)．TheproliferationandsensitivitytoCDDPofMG63／CDDPbeforeandafterapplicationofAs2O3(0．20mg／L)weretestedusingmethylthiazo
6、ltetrazolium(M1Tr)assay．Real—timequantitativepolymerasechainreaction(Real—timePCR)wasemployedtoinvestigatetheexpressionoflet-7，microRNA(miR)一200candsexdeterminingregionY—box2(SOX．2)inMG63／CDDPcellsbe—foreandafterapplicationofAs2O3．Results(1)Inthepresence
7、ofCDDP(3mg／L)，thegrowthrateofMG63／CDDPwassignificantlyhighercomparedwithMG63cells(P<0．01)．ItwasalsodemonstratedthatMG63／CDDPtreatedbyCDDPdevelopedsimilarproliferationtoMG63cellswithouttreatmentofCDDP(P>0．05)．TheICofMG63andMG63／CDDPcellstoCDDPwereabout6．3
8、mg／Land78．9mg／Lre—spectivelyandtheresistanceindexwasabout12．52．f2)TheresistanceofMG63／CDDPcellstoCDDPwasreducedbytreatmentofAs，0(0．20mL)withreversedmultiplesof3．39andrelativereversedeffi—ciencyof76．6％．(3)MG63／CDDPcellsexpr
