PinlsiRNA對(duì)宮頸癌細(xì)胞SiHa增殖和凋亡的影響-論文.pdf

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1、理婦產(chǎn)科進(jìn)展2013年10月第22卷第10期ProgObstetGynecol，Oct．2013，Vo1．22，No．10·論著·PinlsiRNA對(duì)宮頸癌細(xì)胞SiHa增殖和凋亡的影響錢穎，賀曉琪，蔡惠蘭。，董衛(wèi)紅，王澤華(1．同濟(jì)大學(xué)附屬東方醫(yī)院婦產(chǎn)科，上海200120；2．華中科技大學(xué)同濟(jì)醫(yī)學(xué)院附屬協(xié)和醫(yī)院婦產(chǎn)科．武漢430022；3．湖北科技學(xué)院臨床醫(yī)學(xué)院婦產(chǎn)科教研室，成寧437100)【摘要】目的：通過小干擾RNA(siRNA)栽體介導(dǎo)抑制肽基脯氨酰同分異構(gòu)酶(Pin1)表達(dá)，探討Pinl對(duì)宮頸癌細(xì)胞SiHa增殖與凋亡的影響。方法：構(gòu)建pGPU6／

2、GFP／Neo—PinlsiRNA表達(dá)載體，脂質(zhì)體介導(dǎo)將其轉(zhuǎn)染至宮頸癌細(xì)胞SiHa中；實(shí)驗(yàn)設(shè)3組：實(shí)驗(yàn)組(SiHa／pGPU6一PinlsiRNA組)、陰性對(duì)照組(SiHa／pGPU6一con組)和空白對(duì)照組(Si．Ha)。采用RT—PCR、Westernblot法檢測(cè)Pinl的表達(dá)；M1Tr法檢測(cè)細(xì)胞增殖活性；原位末端標(biāo)記法(TUNEL)和流式細(xì)胞術(shù)(FCM)檢測(cè)細(xì)胞凋亡。結(jié)果：轉(zhuǎn)染SiHa／pGPU6．PinlsiRNA的SiHa細(xì)胞中，PinlmRNA和蛋白表達(dá)分別下調(diào)68．1％和54．8％，細(xì)胞增殖顯著抑制。TUNEL顯示典型細(xì)胞凋亡特征，SiHa

3、／pGPU6-PinlsiRNA組的凋亡指數(shù)(AI)為(29．6±14．58)％，顯著高于SiHa／pGPU6一con組l(8．44±5．65)％l和空白對(duì)照組[(5．34±4．47)％】(P

4、靶點(diǎn)?！娟P(guān)鍵詞】RNA干擾；肽基脯氨酰順反異構(gòu)酶；宮頸癌細(xì)胞；增殖；凋亡中圖分類號(hào)：R711文獻(xiàn)標(biāo)志碼：A文章編號(hào)：1004—7379(2013)10—0787—05ImpactofPinlinhibitionbysiRNAonproliferationandapoptosisinhumancervicalcancercelllineSiHa．Qian，Xiaoqi，Huilan，eta1．1．DepartmentofObstetricsandGynecology，EastHospital，TongfiUniversity，Shanghai200120；2

5、．DepartmentofObstetricsandGynecology，UnionHospital，TongjiMedicalCollege，HuazhongUniversityofScienceandTechnology．Wuhan430022：3．DepartmentofObstetricsandGynecology，ClinicafMedicafCol—fege．日MbeUniversityofScienceandTechnology，Xianning437100【Abstract】objective：Toinvestigatetheinhibit

6、oryeffectofPinlsmallinterferenceRNAfsiRNA)ontheexpressionofPinlgeneincervicalcanercellsSiHaandobservetheeffectsofPinlgenesilencingonproliferationandapoptosisinSiHacel1．Methods：AvectorpGPU6／GFP／Neo—PinlfortranscribingsiRNAtargetingPinlgenewasconstructedandintro-ducedintocervicalcan

7、cercellsSiHabyLipofe-ctamine2000．Threegroupsweredesigned：theexperimentalgrouptransfectedwithpGPU6／GFP／Neo—Pinl(SiHa／pGPU6一PinlsiRNA)，thenegativeeontrolgrouptransfectedwithpGPU6／GFP／Neo(SiHa／pGPU6．Con)andthenon—transfectedgroupfSiHa)．ThePinlmRNAandproteinexpressionlevelsofSiHacells

8、werede—tectedbyRT．PCRandWesternbl