大豆异黄酮诱导食管癌细胞凋亡作用研究

马吉祥; 苏军英; 马金姝; 李会庆; 颜云

Abstract

马吉祥,苏军英,马金姝,李会庆,颜云.大豆异黄酮诱导食管癌细胞凋亡作用研究[J].Chinese journal of Epidemiology,2003,24(11):1040-1043

大豆异黄酮诱导食管癌细胞凋亡作用研究

Study on the molecular mechanism of apoptosis in esophageal cancer cells induced by soybean isoflavone

Received:September 16, 2002

DOI：

KeyWord: 大豆异黄酮食管肿瘤细胞凋亡细胞株

English Key Word: Soybean isoflavone Esophageal neoplasms Apoptosis Cell line

FundProject:

Author Name	Affiliation
MA Ji-xiang	Public Health College of Shandong University, Jinan 250012, China
SU Jun-ying	山东省疾病预防控制中心
MA Jin-shu	济南市疾病预防控制中心
LI Hui-qing	山东省医学科学院
YAN Yun	潍坊市人民医院

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Abstract:

目的在体外实验中,探讨大豆异黄酮诱导食管癌细胞发生凋亡的可能性,揭示该凋亡发生与bcl-2和bax之间的关系.方法采用MTT比色法测定大豆异黄酮对食管癌细胞的生长抑制率;以透射电镜和TUNEL 染色法,定性、定量地研究大豆异黄酮与食管癌细胞凋亡的关系;通过免疫组织化学法和逆转录-聚合酶链反应(RT-PCR)检测凋亡相关基因bcl-2和bax 的表达.结果大豆异黄酮对食管癌细胞有明显抑制作用,随大豆异黄酮浓度增加和培养时间的延长而增强;透射电镜可见食管癌细胞出现典型的凋亡细胞形态,如细胞核染色质致密浓缩,或聚集于核膜呈新月形小体;TUNEL染色法可见,经大豆异黄酮处理24、48、72、96 h后,食管癌EC-9706细胞凋亡数明显随时间延长而增加.免疫组织化学法发现经大豆异黄酮处理24、48、72、96 h后,食管癌EC-9706细胞的bcl-2蛋白阳性率减少,bax蛋白阳性率增加.经大豆异黄酮处理24、48、72、96 h后,经RT-PCR检测发现食管癌EC-9706细胞bcl-2和bax的mRNA表达阳性,并显示bcl-2 mRNA条带密度随时间的延长而递减,bax mRNA条带密度随时间的延长而增强.结论诱导食管癌细胞发生凋亡是大豆异黄酮抗食管癌作用的机制之一,大豆异黄酮可能通过下调bcl-2的表达和上调bax的表达而诱导食管癌细胞发生凋亡.

English Abstract:

ObjectiveTo investigate the function of ap optosis in esophageal cancer cells induced by soybean isoflavone,and the relatio n between this apoptosis and expression of bcl-2 and bax. MethodsIn vitro experiments, MTT assay was used to det ermine the cell growth inhibitory rate.Transmission electron microscope and T UNEL staining method were used to quantitatively and qualitively detect the apop tosis status of esophageal cancer cell line EC-9706 before and after the soybea n isoflavone treatment. Immunohistochemical staining and reverse transcription- polymerase chain reaction were used to detect the expression of apoptosis-regul ated gene bcl-2 and bax. ResultsSoybean isoflavone inhibited the growth of esopha geal cancer cell line EC-9706 in a dose - and time-dependent manner. Soybean isoflavone induced EC-9706 cells to undergo apoptosis with typically apopto tic ch aracteristics,including morphological changes of chromatin condensation, chromat in crescent formation,nucleus fragmentation and apoptotic body formation by tran smission electron microscope and staining positive cells,using TUNEL assay. So ybea n isoflavone reduced the expression of apoptosis-regulated gene bcl-2, and im proving the expression of apoptosis-regulated gene bax. ConclusionSoybean isoflavone seemed to be able to induce the apoptosis in esophageal cancer. This type of apoptosis might be mediated by down-expression of apoptosis-regulated gene bcl-2 and up-expression of apop tosis-regulated gene bax.

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