中国肿瘤临床
中國腫瘤臨床
중국종류림상
CHINESE JOURNAL OF CLINICAL ONCOLOGY
2013年
13期
758-762
,共5页
邹少娜%林敏%陈波%罗招阳
鄒少娜%林敏%陳波%囉招暘
추소나%림민%진파%라초양
胃肿瘤%表没食子儿茶素没食子酸酯%凋亡%基因芯片
胃腫瘤%錶沒食子兒茶素沒食子痠酯%凋亡%基因芯片
위종류%표몰식자인다소몰식자산지%조망%기인심편
stomach neoplasms%epigallocatechin-3-gallate%apoptosis%gene array
目的：研究表没食子儿茶素没食子酸酯（epigallocatechin-3-gallate，EGCG）诱导人胃癌 MGC-803细胞凋亡的生物学效应，并探讨其分子机制。方法：运用DNA含量分析、Annexin Ⅴ/PI双标记法、DNA琼脂糖凝胶电泳以及透射电子显微镜形态学方法来观察细胞凋亡。通过SuperArray人细胞凋亡基因芯片检测100μmol/L EGCG作用于MGC-803细胞12 h后凋亡相关基因的表达谱，采用 RT-PCR 和 Western blot 技术验证上调基因 Fas-L 和下调基因 Bag-1。结果：25、50、100、200μmol/L EGCG 作用于MGC-803细胞24 h后，DNA含量分析出现了明显的亚二倍体峰；100μmol/L EGCG作用4、8、12、24 h后，Annexin Ⅴ/PI双标记法表明早期凋亡细胞显著增加。100μmol/L EGCG作用24 h后，在琼脂糖凝胶上电泳可见DNA凋亡特征性的阶梯状条带。电子显微镜观察到细胞体积缩小，核浓缩和凋亡小体形成等典型的形态学改变。通过基因芯片检测发现，100μmol/L EGCG作用12 h后有8个凋亡相关基因表达差异显著，选择其中 Fas-L 和 Bag-1运用 RT-PCR 和 Western blot 技术进行验证，其结果与基因芯片一致。结论：EGCG能够诱导人胃癌MGC-803细胞凋亡，这可能是多个基因和多条信号转导通路共同作用的结果。
目的：研究錶沒食子兒茶素沒食子痠酯（epigallocatechin-3-gallate，EGCG）誘導人胃癌 MGC-803細胞凋亡的生物學效應，併探討其分子機製。方法：運用DNA含量分析、Annexin Ⅴ/PI雙標記法、DNA瓊脂糖凝膠電泳以及透射電子顯微鏡形態學方法來觀察細胞凋亡。通過SuperArray人細胞凋亡基因芯片檢測100μmol/L EGCG作用于MGC-803細胞12 h後凋亡相關基因的錶達譜，採用 RT-PCR 和 Western blot 技術驗證上調基因 Fas-L 和下調基因 Bag-1。結果：25、50、100、200μmol/L EGCG 作用于MGC-803細胞24 h後，DNA含量分析齣現瞭明顯的亞二倍體峰；100μmol/L EGCG作用4、8、12、24 h後，Annexin Ⅴ/PI雙標記法錶明早期凋亡細胞顯著增加。100μmol/L EGCG作用24 h後，在瓊脂糖凝膠上電泳可見DNA凋亡特徵性的階梯狀條帶。電子顯微鏡觀察到細胞體積縮小，覈濃縮和凋亡小體形成等典型的形態學改變。通過基因芯片檢測髮現，100μmol/L EGCG作用12 h後有8箇凋亡相關基因錶達差異顯著，選擇其中 Fas-L 和 Bag-1運用 RT-PCR 和 Western blot 技術進行驗證，其結果與基因芯片一緻。結論：EGCG能夠誘導人胃癌MGC-803細胞凋亡，這可能是多箇基因和多條信號轉導通路共同作用的結果。
목적：연구표몰식자인다소몰식자산지（epigallocatechin-3-gallate，EGCG）유도인위암 MGC-803세포조망적생물학효응，병탐토기분자궤제。방법：운용DNA함량분석、Annexin Ⅴ/PI쌍표기법、DNA경지당응효전영이급투사전자현미경형태학방법래관찰세포조망。통과SuperArray인세포조망기인심편검측100μmol/L EGCG작용우MGC-803세포12 h후조망상관기인적표체보，채용 RT-PCR 화 Western blot 기술험증상조기인 Fas-L 화하조기인 Bag-1。결과：25、50、100、200μmol/L EGCG 작용우MGC-803세포24 h후，DNA함량분석출현료명현적아이배체봉；100μmol/L EGCG작용4、8、12、24 h후，Annexin Ⅴ/PI쌍표기법표명조기조망세포현저증가。100μmol/L EGCG작용24 h후，재경지당응효상전영가견DNA조망특정성적계제상조대。전자현미경관찰도세포체적축소，핵농축화조망소체형성등전형적형태학개변。통과기인심편검측발현，100μmol/L EGCG작용12 h후유8개조망상관기인표체차이현저，선택기중 Fas-L 화 Bag-1운용 RT-PCR 화 Western blot 기술진행험증，기결과여기인심편일치。결론：EGCG능구유도인위암MGC-803세포조망，저가능시다개기인화다조신호전도통로공동작용적결과。
Objective: This study investigates the biological effects and explores the molecular mechanisms of epigallocate-chin-3-gallate (EGCG) on the apoptosis of the human gastric cancer MGC-803 cells. Methods: After treatment with EGCG, cell apopto-sis was verified by flow cytometry with Annexin V and propidium iodide staining, DNA agarose gel electrophoresis, and transmission electron microscopy. The expression profiles of the apoptosis-related genes in the MGC-803 cells with or without treatment by EGCG for 12 h (100 μmol/L), was identified using SuperArray Human Apoptosis Gene Array. The upregulated Fas-L gene and down-regulated Bag-1 gene were confirmed by reverse transcription polymerase chain reaction (RT-PCR) and Western blot. Results: When the MGC-803 cells were treated with EGCG at 25, 50, 100, and 200 μmol/L for 24 h, evident sub-diploid peaks were observed. Under treat-ment with 100 μmol/L for 4, 8, 12, and 24 h, the number of early apoptotic cells was greatly increased. When the cells were treated with 100 μmol/L for 24 h, the DNA extracted from the cells displayed a characteristic ladder pattern with agarose gel electrophoresis. Typi-cal morphological changes were observed by electron microscopy, including cell shrinkage, karyo-pyknosis, and the formation of apop-totic bodies. The differential expressions of eight apoptosis-associated genes were determined by gene array detection. The results of Fas-L and Bag-1 selected for RT-PCR and Western blot were consistent with those of gene array. Conclusion: EGCG induces apoptosis in MGC-803 cells, which might be mediated by a number of specific genes and various signal transduction pathways.
