中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2008年
7期
493-497
,共5页
代志军%高洁%王西京%纪宗正%董济民%刘小旭%管海涛%康华峰%刁岩
代誌軍%高潔%王西京%紀宗正%董濟民%劉小旭%管海濤%康華峰%刁巖
대지군%고길%왕서경%기종정%동제민%류소욱%관해도%강화봉%조암
胰腺肿瘤%乏氧%CoCl2%HIF-1α%细胞凋亡
胰腺腫瘤%乏氧%CoCl2%HIF-1α%細胞凋亡
이선종류%핍양%CoCl2%HIF-1α%세포조망
Pancreatic neoplasms%Hypoxia%Cobalt chloride%HIF-1α%Apoptosis
目的 探讨CoCl2诱导缺氧对体外培养的胰腺癌PC-2细胞缺氧诱导因子-1(HIF-1α)表达变化及诱导凋亡作用.方法 以50、100、150、200μmol/L不同浓度CoCl2处理PC-2细胞模拟肿瘤体内乏氧模型,MTT法检测不同浓度、不同时间CoCl2处理后的PC-2细胞生长情况;透射电镜下观察乏氧微环境作用下的PC-2细胞超微结构的改变;免疫细胞化学和RT-PCR方法 检测乏氧微环境对PC-2细胞HIF-1α基因表达的影响;AnnexinV-FITC/PI双标记染色,流式细胞术检测肿瘤细胞凋亡情况.结果 不同浓度CoCl2对胰腺癌PC-2细胞生长曲线有不同程度的改变,对数生长期和平台期提前,对数期缩短.随着CoCl2浓度增加,曲线越趋于低平.透射电镜下可见线粒体肿胀,染色质边集等凋亡早期表现,少见凋亡小体.免疫细胞化学和RT-PCR结果 显示,化学缺氧剂COCl2模拟的乏氧微环境可引起胰腺癌PC-2细胞HIF-1α表达的上调;流式细胞仪检测结果 显示,正常对照组、100、150、200 μmol/L CoCl2不同浓度组PC-2细胞凋亡率分别为10.77%、34.32%、40.17%和52.30%.结论 CoCl2对胰腺癌PC-2细胞具有一定的凋亡诱导作用,同时也缩短了细胞正常的生长期,细胞形态发生相应改变,可能与上调HIF-1α的表达有关.
目的 探討CoCl2誘導缺氧對體外培養的胰腺癌PC-2細胞缺氧誘導因子-1(HIF-1α)錶達變化及誘導凋亡作用.方法 以50、100、150、200μmol/L不同濃度CoCl2處理PC-2細胞模擬腫瘤體內乏氧模型,MTT法檢測不同濃度、不同時間CoCl2處理後的PC-2細胞生長情況;透射電鏡下觀察乏氧微環境作用下的PC-2細胞超微結構的改變;免疫細胞化學和RT-PCR方法 檢測乏氧微環境對PC-2細胞HIF-1α基因錶達的影響;AnnexinV-FITC/PI雙標記染色,流式細胞術檢測腫瘤細胞凋亡情況.結果 不同濃度CoCl2對胰腺癌PC-2細胞生長麯線有不同程度的改變,對數生長期和平檯期提前,對數期縮短.隨著CoCl2濃度增加,麯線越趨于低平.透射電鏡下可見線粒體腫脹,染色質邊集等凋亡早期錶現,少見凋亡小體.免疫細胞化學和RT-PCR結果 顯示,化學缺氧劑COCl2模擬的乏氧微環境可引起胰腺癌PC-2細胞HIF-1α錶達的上調;流式細胞儀檢測結果 顯示,正常對照組、100、150、200 μmol/L CoCl2不同濃度組PC-2細胞凋亡率分彆為10.77%、34.32%、40.17%和52.30%.結論 CoCl2對胰腺癌PC-2細胞具有一定的凋亡誘導作用,同時也縮短瞭細胞正常的生長期,細胞形態髮生相應改變,可能與上調HIF-1α的錶達有關.
목적 탐토CoCl2유도결양대체외배양적이선암PC-2세포결양유도인자-1(HIF-1α)표체변화급유도조망작용.방법 이50、100、150、200μmol/L불동농도CoCl2처리PC-2세포모의종류체내핍양모형,MTT법검측불동농도、불동시간CoCl2처리후적PC-2세포생장정황;투사전경하관찰핍양미배경작용하적PC-2세포초미결구적개변;면역세포화학화RT-PCR방법 검측핍양미배경대PC-2세포HIF-1α기인표체적영향;AnnexinV-FITC/PI쌍표기염색,류식세포술검측종류세포조망정황.결과 불동농도CoCl2대이선암PC-2세포생장곡선유불동정도적개변,대수생장기화평태기제전,대수기축단.수착CoCl2농도증가,곡선월추우저평.투사전경하가견선립체종창,염색질변집등조망조기표현,소견조망소체.면역세포화학화RT-PCR결과 현시,화학결양제COCl2모의적핍양미배경가인기이선암PC-2세포HIF-1α표체적상조;류식세포의검측결과 현시,정상대조조、100、150、200 μmol/L CoCl2불동농도조PC-2세포조망솔분별위10.77%、34.32%、40.17%화52.30%.결론 CoCl2대이선암PC-2세포구유일정적조망유도작용,동시야축단료세포정상적생장기,세포형태발생상응개변,가능여상조HIF-1α적표체유관.
Objective To investigate the variation of hypoxia inducible factor-1α (HIF-1α) ex-pression and inducing apoptosis effect of pancreatic cancer PC-2 cells by cobalt chloride (CoCl2) simu-lated hypoxia. Methods MTT assay was used to examine the proliferation of PC-2 cells treated by dif-ferent concentrations of CoCl2 for 24-96 h. The cellular morphology of PC-2 cells was observed by light inverted microscope and transmission electron microscope (EM). The expression of HIF-1α was determined by semi-quantitive RT-PCR and immunohistochemieal SP methods. Flow cytometry was performed to analyze the apoptosis of PC-2 cells by staining with annexinV-FITC/PI. Results The in-hibition of proliferation of PC-2 cells in vitro was observed in time-dependent but no dose-dependent manner. Log phase growth and platform phase advanced. The cellular growth period became shorter and cellular morphology was changed correspondingly. Chondrosomes were swelling,chromatin accu-mulated under the nuclear membrane and apoptosis bodies were also seen by EM. CoCl2 simulated hy-poxia could effectively up-regulate the expression of HIF-1α both at the Mrna and protein levels. The expression of HIF-1α was inhibited in a dose-and time-dependent manner. The apoptotic rate of control group and 100,150,200 μmol/L CoCl2 groups were 10.77%,34.32%,40. 17% and 52.30%, respec-tively. Conclusion Hypoxia simulated by CoCl2 can effectively induce apoptosis in PC-2 cells and cellu-lar growth period becomes shorter. The cellular morphology is changed correspondingly. The mecha-nism might be related to over-expression of HIF-1α.
