中国医师杂志
中國醫師雜誌
중국의사잡지
JOURNAL OF CHINESE PHYSICIAN
2008年
9期
1156-1159
,共4页
张士龙%曾甫清%董继华%朱朝辉%廖贵益%彭世波
張士龍%曾甫清%董繼華%硃朝輝%廖貴益%彭世波
장사룡%증보청%동계화%주조휘%료귀익%팽세파
DNA(胞嘧啶-5-)-甲基转移酶%RNA干扰%膀胱肿瘤
DNA(胞嘧啶-5-)-甲基轉移酶%RNA榦擾%膀胱腫瘤
DNA(포밀정-5-)-갑기전이매%RNA간우%방광종류
DNA(cytosine-5-)-methyhransferase%RNA interference%Bladder neoplasms
目的 体外研究重组质粒pshRNA-DNMT3b对膀胱癌T24细胞中DNMT3bmRNA和蛋白的沉默效应以及细胞增殖抑制的影响,初步探讨DNMT3b在膀胱肿瘤发生过程中的作用.方法 实验分为空白对照组、HK组及pshRNA.DNMT3b组(24、48、72 h);常规培养T24细胞,用脂质体lipofectamine2000将重组质粒pshRNA-DNMT3b转染进入T24细胞,后进行RT-PCR、Western Blot、MTT检测,观察pshRNA-DNMT3b对T24细胞中DNMT3bmRNA、蛋白质水平变化以及细胞增殖的影响.结果 重组质粒pshRNA-DNMT3b成功的转染进入T24细胞;RT-PCR电泳结果用Gel-pro analyzer图像分析系统进行灰度分析,空白对照组、HK组、pshRNA-DNMT3b组(24、48、72 h)条带的IOD相对比值(DNMT3/13.actin)分别是(99.56±1.24)%、(99.12±1.35)%、(75.77±1.42)%、(44.69±1.05)%和(20.52±0.89)%;Westem blot图像分析结果各组的相对比值分别是(99.43±1.28)%、(98.90±1.31)%、(67.83±1.02)%、(43.43±1.05)%和(21.92±0.89)%;在MTT检测中空白对照组和HK组之间差异无统计学意义(P＞0.05),pshRNA-DNMT3b组仅在72 h后和前2组之间差异有统计学意义(P＜0.01),但抑制率仅增加0.45%左右.结论 重组质粒pshRNA-DNMT3b能有效的降低膀胱癌T24细胞中基因DNMT3b mRNA的转录及蛋白的表达,但在抑制细胞增殖上作用甚微.
目的 體外研究重組質粒pshRNA-DNMT3b對膀胱癌T24細胞中DNMT3bmRNA和蛋白的沉默效應以及細胞增殖抑製的影響,初步探討DNMT3b在膀胱腫瘤髮生過程中的作用.方法 實驗分為空白對照組、HK組及pshRNA.DNMT3b組(24、48、72 h);常規培養T24細胞,用脂質體lipofectamine2000將重組質粒pshRNA-DNMT3b轉染進入T24細胞,後進行RT-PCR、Western Blot、MTT檢測,觀察pshRNA-DNMT3b對T24細胞中DNMT3bmRNA、蛋白質水平變化以及細胞增殖的影響.結果 重組質粒pshRNA-DNMT3b成功的轉染進入T24細胞;RT-PCR電泳結果用Gel-pro analyzer圖像分析繫統進行灰度分析,空白對照組、HK組、pshRNA-DNMT3b組(24、48、72 h)條帶的IOD相對比值(DNMT3/13.actin)分彆是(99.56±1.24)%、(99.12±1.35)%、(75.77±1.42)%、(44.69±1.05)%和(20.52±0.89)%;Westem blot圖像分析結果各組的相對比值分彆是(99.43±1.28)%、(98.90±1.31)%、(67.83±1.02)%、(43.43±1.05)%和(21.92±0.89)%;在MTT檢測中空白對照組和HK組之間差異無統計學意義(P＞0.05),pshRNA-DNMT3b組僅在72 h後和前2組之間差異有統計學意義(P＜0.01),但抑製率僅增加0.45%左右.結論 重組質粒pshRNA-DNMT3b能有效的降低膀胱癌T24細胞中基因DNMT3b mRNA的轉錄及蛋白的錶達,但在抑製細胞增殖上作用甚微.
목적 체외연구중조질립pshRNA-DNMT3b대방광암T24세포중DNMT3bmRNA화단백적침묵효응이급세포증식억제적영향,초보탐토DNMT3b재방광종류발생과정중적작용.방법 실험분위공백대조조、HK조급pshRNA.DNMT3b조(24、48、72 h);상규배양T24세포,용지질체lipofectamine2000장중조질립pshRNA-DNMT3b전염진입T24세포,후진행RT-PCR、Western Blot、MTT검측,관찰pshRNA-DNMT3b대T24세포중DNMT3bmRNA、단백질수평변화이급세포증식적영향.결과 중조질립pshRNA-DNMT3b성공적전염진입T24세포;RT-PCR전영결과용Gel-pro analyzer도상분석계통진행회도분석,공백대조조、HK조、pshRNA-DNMT3b조(24、48、72 h)조대적IOD상대비치(DNMT3/13.actin)분별시(99.56±1.24)%、(99.12±1.35)%、(75.77±1.42)%、(44.69±1.05)%화(20.52±0.89)%;Westem blot도상분석결과각조적상대비치분별시(99.43±1.28)%、(98.90±1.31)%、(67.83±1.02)%、(43.43±1.05)%화(21.92±0.89)%;재MTT검측중공백대조조화HK조지간차이무통계학의의(P＞0.05),pshRNA-DNMT3b조부재72 h후화전2조지간차이유통계학의의(P＜0.01),단억제솔부증가0.45%좌우.결론 중조질립pshRNA-DNMT3b능유효적강저방광암T24세포중기인DNMT3b mRNA적전록급단백적표체,단재억제세포증식상작용심미.
Objective To investigate the effect of recombinant plasmid pshRNA-DNMT3b on expression of DNMT3b mRNA and protein and on the proliferation of bladder cancer T24 cells,and research the function of DNMT3b in the process of bladder tumor formation.Methods There were three groups in this study,which are blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),respectively.T24 cells were cultured routinely and transfected by the recombinant plasmids with lipfectamine 2000.The cells were detected by methods of RT-PCR,western blot and MTT.The varying level of DNMT3b mRNA and expression protein,and the conditions of cellular survival rate were observed.Results The recombinant plasmids were successfully transfected into T24 cell lines.The grey valHe of RT-PCR elctrophoretogram was analyzed by the software of Gel-pro analyzer,the rate of blank controller,HK and pshRNA-DNMT3b(24h,48h,72h),was (99.56±1.24)%,(99.12±1.35)%,(75.77±1.42)%,(44.69±1.05)%and(20.52±0.89)%,respectively.The analytical resuit of western blot image was(99.43±1.28)%,(98.90±1.31)%,(67.83±1.02)%,(43.43±1.05)%and(21.92±0.89)%.There was no statistically difference in survival between blank control and HK(P＞0.05).The group of pshRNA-DNMT3b and other two groups had statistical difference only at the 72th hour and the cell inhibitory growth rate only increase 0.45%.Conclusions The recombinant ptasmid pshRNA-DNMT3b can inhibit the expression of mRNA and protein of DNMT3b effectively.However,it has slight function on inhibiting cell proliferation.
