il18基因轉(zhuǎn)染對小鼠卵巢癌ovhm體外增殖及體內(nèi)成瘤的影響

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1、IL18基因轉(zhuǎn)染對小鼠卵巢癌OVHM體外增殖及體內(nèi)成瘤的影響作者：崔澂,郝淑維,李新偉,白小嘉,程建新,單保恩【摘要】　　目的:分析轉(zhuǎn)染IL18基因后,對小鼠卵巢癌OVHM細(xì)胞體外增殖、凋亡及體內(nèi)成瘤的影響,初步探討IL18基因轉(zhuǎn)染治療卵巢癌的應(yīng)用價值。方法:將逆轉(zhuǎn)錄病毒攜帶的小鼠IL18基因成功轉(zhuǎn)染至OVHM（OVHM/IL18）,以空載體轉(zhuǎn)染的OVHM（OVHM/LXSN）和野生型（OVHM）作為對照。分別采用MTT、FCM檢測體外培養(yǎng)細(xì)胞的增殖、凋亡及周期分布。于裸鼠皮下分別接種細(xì)胞,觀察各組種植瘤生長情況,計算成瘤率;RTPCR檢測瘤組織中IL18mRNA表達(dá);FCM

2、和電鏡分析腫瘤細(xì)胞增殖、凋亡狀況。結(jié)果:3組細(xì)胞的體外增殖未見明顯差異,均無明顯凋亡現(xiàn)象,增殖指數(shù)、細(xì)胞周期分布均無明顯差異（均P>0.05）。接種后,3組裸鼠的成瘤率相同,但OVHM/IL18組成瘤時間較晚,隨瘤齡增長腫瘤生長緩慢,瘤組織中IL18mRNA陽性表達(dá)。OVHM/IL18組裸鼠種植瘤細(xì)胞可見典型的凋亡現(xiàn)象,G0/G1期細(xì)胞明顯增多,S期細(xì)胞明顯減少,增殖指數(shù)明顯降低,凋亡指數(shù)明顯增高（P<0.01）。結(jié)論:IL18基因成功轉(zhuǎn)染后,雖對體外卵巢癌細(xì)胞無直接毒性,但可能在體內(nèi)借助非直接殺傷的其他途徑,通過阻斷細(xì)胞周期、促進(jìn)腫瘤細(xì)胞凋亡等抑制體內(nèi)成瘤。15【關(guān)鍵

3、詞】IL18基因轉(zhuǎn)染OVHM增殖凋亡成瘤　　［Abstract］AIM:ToinvestigatetheeffectsofIL18genetransfectiononproliferation,apoptosisandtumorigenesisofmouseovariancancercelllineOVHM.METHODS:OVHMweretransfectedwithretrovirusencodingIL18gene(OVHM/IL18).ThewildOVHMandOVHMtransfectedwithvectorwithoutIL18gene(OVHM/LXSN)weree

4、nrolledasthecontrols.Theproliferation,cellcycleandapoptosisweremeasuredbyMTTandFCMrespectively.Thenudemiceweresubcutaneouslyinoculatedwiththethreedifferentcellsrespectively.Theobservationoftransplantedtumor’sgrowthandcalculationoftumorigenesisratiowasmade.TheexpressionofIL18mRNAintumortissueswasd

5、etectedbyRTPCR,andtheapoptosisoftumorcellintissuewasanalyzedbyelectronmicroscopeandFCM.RESULTS:Thethreecellsculturedinvitroshowednoapoptoticpeaks,andnodifferencesinproliferationandcellcycle(AllP>0.05).Followinginoculation,theratiosoftumorigenesisweresimilarinallthethreegroups.ButinOVHM/IL18gr

6、oup,thelatentperiodoftumorigenesiswaslongerwithslowertumorgrowthrateandpositiveexpressionofIL18mRNAintumortissue.Itwasalsoobservedthatinthetumorcellsfromnudemiceinoculated15byOVHM/IL18,thecellsinphaseofG0/G1increasedwithtypicalmorphologyofapoptosis,andthoseinSphasedecreasedwithdecreasedprolifera

7、tionindexandincreasedapoptosisindex(AllP<0.01).CONCLUSION:AlthoughIL18genehasnocytotoxiceffectsinvitro,itsinhibitingeffectsontumorigenesisofOVHMinvivowereexertedbyinterdictingcellcycleandacceleratingapoptosis