中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2015年
2期
129-132
,共4页
任扩军%戚晓东%宋瑾%王静
任擴軍%慼曉東%宋瑾%王靜
임확군%척효동%송근%왕정
乳腺肿瘤%受体,雌激素%细胞凋亡%基因疗法
乳腺腫瘤%受體,雌激素%細胞凋亡%基因療法
유선종류%수체,자격소%세포조망%기인요법
Breast neoplasms%Receptors,estrogen%Apoptosis%Gene therapy
目的 探讨重组人p53腺病毒(Ad-p53)联合氟维司群对人乳腺癌细胞MCF-7凋亡的影响.方法 采用雌激素受体(ER)阳性MCF-7乳腺癌细胞为研究对象,设对照组(空载病毒组)、氟维司群组、Ad-p53组、Ad-p53+氟维司群组作用于MCF-7细胞,处理72 h后,倒置显微镜下观察各组细胞的形态学变化;采用蛋白质印迹法(Western blot)检测各组细胞p53蛋白、ER蛋白的表达水平;流式细胞术法观察细胞的凋亡情况;MTS法观察MCF-7各组细胞增殖抑制的情况.结果 Ad-p53与氟维司群联合作用于MCF-7细胞后,细胞生长受到显著抑制,几乎无增长;流式细胞术观察到单用Ad-p53和氟维司群细胞凋亡率分别是(14.5±2.9)％、(13.3±1.2)％,联合用药组细胞凋亡率为(38.1±5.9)％,显著高于单用药组(P =0.001);MTS法得到Ad-p53和氟维司群分别处理MCF-7细胞24、48、72、96 h后增殖抑制率分别为(8.21±0.54)％、(28.50±1.42)％、(50.14±0.78)％、(58.25±2.92)％和(9.73±1.68)％、(25.26±0.82)％、(35.25±3.94)％、(46.37±2.56)％,联合用药处理24、48、72、96 h后,细胞的增殖抑制率分别为(12.42±1.76)％、(35.20±0.58)％、(62.08±2.56)％及(75.43±3.56)％,联合用药组细胞增殖抑制率在各个时间段均高于单用Ad-p53或氟维司群组(P＜0.05);蛋白质印迹检测表明对照组和氟维司群组p53蛋白低表达,Ad-p53组和联合用药组p53蛋白高表达,但是组间差异无统计学意义;Ad-p53组ER蛋白表达高于对照组,联合用药组ER蛋白表达明显低于单用Ad-p53组(P＜0.05).结论 氟维司群联合重组人p53腺病毒可显著促进乳腺癌细胞凋亡,抑制细胞增殖,二者可能具有协同作用.
目的 探討重組人p53腺病毒(Ad-p53)聯閤氟維司群對人乳腺癌細胞MCF-7凋亡的影響.方法 採用雌激素受體(ER)暘性MCF-7乳腺癌細胞為研究對象,設對照組(空載病毒組)、氟維司群組、Ad-p53組、Ad-p53+氟維司群組作用于MCF-7細胞,處理72 h後,倒置顯微鏡下觀察各組細胞的形態學變化;採用蛋白質印跡法(Western blot)檢測各組細胞p53蛋白、ER蛋白的錶達水平;流式細胞術法觀察細胞的凋亡情況;MTS法觀察MCF-7各組細胞增殖抑製的情況.結果 Ad-p53與氟維司群聯閤作用于MCF-7細胞後,細胞生長受到顯著抑製,幾乎無增長;流式細胞術觀察到單用Ad-p53和氟維司群細胞凋亡率分彆是(14.5±2.9)％、(13.3±1.2)％,聯閤用藥組細胞凋亡率為(38.1±5.9)％,顯著高于單用藥組(P =0.001);MTS法得到Ad-p53和氟維司群分彆處理MCF-7細胞24、48、72、96 h後增殖抑製率分彆為(8.21±0.54)％、(28.50±1.42)％、(50.14±0.78)％、(58.25±2.92)％和(9.73±1.68)％、(25.26±0.82)％、(35.25±3.94)％、(46.37±2.56)％,聯閤用藥處理24、48、72、96 h後,細胞的增殖抑製率分彆為(12.42±1.76)％、(35.20±0.58)％、(62.08±2.56)％及(75.43±3.56)％,聯閤用藥組細胞增殖抑製率在各箇時間段均高于單用Ad-p53或氟維司群組(P＜0.05);蛋白質印跡檢測錶明對照組和氟維司群組p53蛋白低錶達,Ad-p53組和聯閤用藥組p53蛋白高錶達,但是組間差異無統計學意義;Ad-p53組ER蛋白錶達高于對照組,聯閤用藥組ER蛋白錶達明顯低于單用Ad-p53組(P＜0.05).結論 氟維司群聯閤重組人p53腺病毒可顯著促進乳腺癌細胞凋亡,抑製細胞增殖,二者可能具有協同作用.
목적 탐토중조인p53선병독(Ad-p53)연합불유사군대인유선암세포MCF-7조망적영향.방법 채용자격소수체(ER)양성MCF-7유선암세포위연구대상,설대조조(공재병독조)、불유사군조、Ad-p53조、Ad-p53+불유사군조작용우MCF-7세포,처리72 h후,도치현미경하관찰각조세포적형태학변화;채용단백질인적법(Western blot)검측각조세포p53단백、ER단백적표체수평;류식세포술법관찰세포적조망정황;MTS법관찰MCF-7각조세포증식억제적정황.결과 Ad-p53여불유사군연합작용우MCF-7세포후,세포생장수도현저억제,궤호무증장;류식세포술관찰도단용Ad-p53화불유사군세포조망솔분별시(14.5±2.9)％、(13.3±1.2)％,연합용약조세포조망솔위(38.1±5.9)％,현저고우단용약조(P =0.001);MTS법득도Ad-p53화불유사군분별처리MCF-7세포24、48、72、96 h후증식억제솔분별위(8.21±0.54)％、(28.50±1.42)％、(50.14±0.78)％、(58.25±2.92)％화(9.73±1.68)％、(25.26±0.82)％、(35.25±3.94)％、(46.37±2.56)％,연합용약처리24、48、72、96 h후,세포적증식억제솔분별위(12.42±1.76)％、(35.20±0.58)％、(62.08±2.56)％급(75.43±3.56)％,연합용약조세포증식억제솔재각개시간단균고우단용Ad-p53혹불유사군조(P＜0.05);단백질인적검측표명대조조화불유사군조p53단백저표체,Ad-p53조화연합용약조p53단백고표체,단시조간차이무통계학의의;Ad-p53조ER단백표체고우대조조,연합용약조ER단백표체명현저우단용Ad-p53조(P＜0.05).결론 불유사군연합중조인p53선병독가현저촉진유선암세포조망,억제세포증식,이자가능구유협동작용.
Objective To explore the apoptosis-inducing effect of Ad-p53 plus fulvestrant in MCF-7 breast cancer cells.Methods ER-positive MCF-7 breast cancer cells were used.MCF-7 cells were established with control,fulvestrant alone,Ad-p53 alone and Ad-p53 plus fulvestrant.The morphological changes of MCF-7 after drug combination were observed by inverted microscopy.Methyltetrazolium salt (MTS) assay was used to determine the proliferation of MCF-7 cells.The protein expressions of p53 and ER were examined by Western blot.Flow cytometry was used to detect apoptosis.Results Significant inhibition of cell growth was observed by inverted microscopy.Flow cytometry revealed that the apoptosis rate of using fulvestrant alone or Ad-p53 alone was(13.3 ± 1.2)％ and(14.5 ± 2.9)％.The apoptosis rate of combined group was (38.1 ± 5.9) ％ and it was higher than Ad-p53 or fulvestrant alone (P =0.001,P =0.001).MTS assay indicated that,after treating for 24,48,72,96 h by Ad-p53 alone or fulvestrant alone,the inhibition of cell proliferation rate was (8.21 ± 0.54) ％,(28.50 ± 1.42) ％,(50.14 ± 0.78) ％,(58.25 ±2.92)％ and (9.73 ± 1.68)％,(25.26 ±0.82)％,(35.25 ±3.94)％ and (46.37 ±2.56)％ respectively.Simultaneously,the inhibition of cell proliferation rate of combined group was (12.42 ± 1.76)％,(35.20 ±0.58)％,(62.08 ±2.56)％ and (75.43 ±3.56)％.It further confirmed the increase of cell proliferation inhibition rate after drug combination (P ＜ 0.05).Western blot showed that the p53 protein expression level increased obviously with Ad-p53 in MCF-7cells.Ad-p53 increased the level of ER expression while drug combination reduced the level of ER expression.Conclusions Ad-p53 plus fulvestrant can significantly inhibit cell growth and induce cell apoptosis.And there is probably some synergistic effects between both.
