中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
4期
718-721
,共4页
俞洋%宗祥云%冯建国%吴斌
俞洋%宗祥雲%馮建國%吳斌
유양%종상운%풍건국%오빈
胶质母细胞瘤%血管内皮生长因子%RNA干扰%放射治疗%化学治疗
膠質母細胞瘤%血管內皮生長因子%RNA榦擾%放射治療%化學治療
효질모세포류%혈관내피생장인자%RNA간우%방사치료%화학치료
Glioblastoma%Vascular endothelial growth factor%RNA interference%Radiotherapy%Chemotherapy
目的 探讨血管内皮生长因子(VEGF) mRNA干扰对胶质母细胞瘤的治疗意义.方法 应用RNA干扰技术,对胶质母细胞瘤细胞株U251进行了VEGF mRNA干扰、辅以6Gy的放射剂量照射、化疗处理.在干扰前后用流式细胞技术、噻唑蓝(MTT)比色法及倒置显微镜观察等技术对U251细胞株的细胞周期、凋亡率、抑制率、细胞形态进行检测.结果 胶质母细胞瘤U251细胞在转染VEGF小干扰RNA(siRNA)后,逆转录-聚合酶链反应(RT-PCR)检测发现VEGF mRNA的表达显著下降,均数下调大于60％；流式细胞技术显示,U251转染VEGFsiRNA后细胞出现G0～G1阻滞,G2 +M期细胞数目减少,转染细胞出现不同程度凋亡；MTT比色法可见,转染前不同浓度的紫杉醇对U251细胞有一定的抑制,作用48 h半数抑制剂量(IC50)=28.1 g/L.干扰后,紫杉醇在不同浓度对U251细胞的抑制率都有大幅度的提高,IC50=0.02 g/L；集落形成抑制实验中,单药物组与单放疗组在克隆形成率上差异无统计学意义(P＞0.05),U251细胞转染VEGFsiRNA后,明显抑制肿瘤细胞的集落形成,并对紫杉醇及放射治疗具有明显的协同作用；细胞学形态变化观察发现,转染后实验组的细胞形态明显差于未转染组细胞.结论 VEGF基因干扰可以抑制U251细胞的增殖、促进其凋亡,并能提高U251细胞对放化疗的敏感性.脂质体紫杉醇在细胞实验中表现出较好的抑制肿瘤作用及对VEGF基因干扰和放疗的协同作用.
目的 探討血管內皮生長因子(VEGF) mRNA榦擾對膠質母細胞瘤的治療意義.方法 應用RNA榦擾技術,對膠質母細胞瘤細胞株U251進行瞭VEGF mRNA榦擾、輔以6Gy的放射劑量照射、化療處理.在榦擾前後用流式細胞技術、噻唑藍(MTT)比色法及倒置顯微鏡觀察等技術對U251細胞株的細胞週期、凋亡率、抑製率、細胞形態進行檢測.結果 膠質母細胞瘤U251細胞在轉染VEGF小榦擾RNA(siRNA)後,逆轉錄-聚閤酶鏈反應(RT-PCR)檢測髮現VEGF mRNA的錶達顯著下降,均數下調大于60％；流式細胞技術顯示,U251轉染VEGFsiRNA後細胞齣現G0～G1阻滯,G2 +M期細胞數目減少,轉染細胞齣現不同程度凋亡；MTT比色法可見,轉染前不同濃度的紫杉醇對U251細胞有一定的抑製,作用48 h半數抑製劑量(IC50)=28.1 g/L.榦擾後,紫杉醇在不同濃度對U251細胞的抑製率都有大幅度的提高,IC50=0.02 g/L；集落形成抑製實驗中,單藥物組與單放療組在剋隆形成率上差異無統計學意義(P＞0.05),U251細胞轉染VEGFsiRNA後,明顯抑製腫瘤細胞的集落形成,併對紫杉醇及放射治療具有明顯的協同作用；細胞學形態變化觀察髮現,轉染後實驗組的細胞形態明顯差于未轉染組細胞.結論 VEGF基因榦擾可以抑製U251細胞的增殖、促進其凋亡,併能提高U251細胞對放化療的敏感性.脂質體紫杉醇在細胞實驗中錶現齣較好的抑製腫瘤作用及對VEGF基因榦擾和放療的協同作用.
목적 탐토혈관내피생장인자(VEGF) mRNA간우대효질모세포류적치료의의.방법 응용RNA간우기술,대효질모세포류세포주U251진행료VEGF mRNA간우、보이6Gy적방사제량조사、화료처리.재간우전후용류식세포기술、새서람(MTT)비색법급도치현미경관찰등기술대U251세포주적세포주기、조망솔、억제솔、세포형태진행검측.결과 효질모세포류U251세포재전염VEGF소간우RNA(siRNA)후,역전록-취합매련반응(RT-PCR)검측발현VEGF mRNA적표체현저하강,균수하조대우60％；류식세포기술현시,U251전염VEGFsiRNA후세포출현G0～G1조체,G2 +M기세포수목감소,전염세포출현불동정도조망；MTT비색법가견,전염전불동농도적자삼순대U251세포유일정적억제,작용48 h반수억제제량(IC50)=28.1 g/L.간우후,자삼순재불동농도대U251세포적억제솔도유대폭도적제고,IC50=0.02 g/L；집락형성억제실험중,단약물조여단방료조재극륭형성솔상차이무통계학의의(P＞0.05),U251세포전염VEGFsiRNA후,명현억제종류세포적집락형성,병대자삼순급방사치료구유명현적협동작용；세포학형태변화관찰발현,전염후실험조적세포형태명현차우미전염조세포.결론 VEGF기인간우가이억제U251세포적증식、촉진기조망,병능제고U251세포대방화료적민감성.지질체자삼순재세포실험중표현출교호적억제종류작용급대VEGF기인간우화방료적협동작용.
Objective To explore the effect of vascular endothelial growth factor (VEGF) mRNA interference on glioblatoma.Methods After knockdown of VEGF mRNA expression using VEGF short hairpin RNA (shRNA),glioma U251 cell were treated with chemotherapy or radiotherapy or chemotherapy plus radiotherapy or without any treatment.The changes in cell cycle,apoptosis rate,cell colony-formation and cell morphology were observed.Results After knockdown of VEGF mRNA expression using VEGF shRNA,VEGF mRNA expression levels in glioma U251 cells were inhibited significantly (P ＜ 0.001).The flow cytometry revealed that both control cells and VEGF shRNA-transfected cells exhibited G0-G1 arrest,and reduced number of cells in the G2 and M phases.The apoptosis rate of VEGF shRNA-transfected cells was increased as compared with the control cells.It was found that various concentrations of paciltaxel reduced U251 cell viability 50％ inhibitory dose(IC50) =28.1 mg/L,and VEGF knockdown significantly sensitized U251 cells to paclitaxel (IC50 =0.02 mg/L).Groups with drug treatment alone and the radiotherapy alone showed no significant difference (P ＞ 0.05).After VEGF knockdown,colony formation in paclitaxel and radiation treated U251 cells was significantly reduced (17.57％ to 6.33％,P ＜ 0.01).Under the microscopy,it was found VEGF shRNA-transfected cells showed worse cellular morphology than non-transfected cells.Conclusion The interference of VEGF gene expression can inhibit the proliferation of U251cells,promote apoptosis of U251 cells,and increase the sensitivity of U251 cells to chemotherapy and radiotherapy.Liposomal paclitaxel can enhance drug delivery in vivo,although the VEGF gene interference and the practical application of liposomal paclitaxel remains to be tested and explored in future studies.
