桦褐孔菌醇提物对胃癌侵袭迁移的影响及分子机制探讨  

Effect of alcohol extract of Inonotus obliquus on invasion and migration of gastric carcinoma and the exploration of its molecular mechanism

作　　者：王蔚 乔羽 周忠光 杨婧[4] 刘旭[5] 仲丽丽 WANG Wei;QIAO Yu;ZHOU Zhong-guang(Teaching and Researching Section of Medical Examination,College of Medical Examination,Liaoning University of Traditional Chinese Medicine,Shenyang 110000,China;不详)

机构地区：[1]辽宁中医药大学医学检验学院医学检验教研室,辽宁沈阳110000 [2]黑龙江中医药大学基础医学院伤寒教研室,黑龙江哈尔滨150000 [3]黑龙江中医药大学基础医学院组织胚胎学教研室,黑龙江哈尔滨150000 [4]黑龙江中医药大学基础医学院病理教研室,黑龙江哈尔滨150000 [5]黑龙江中医药大学实验中心黑龙江,哈尔滨150000 [6]黑龙江中医药大学临床医学院病理科,黑龙江哈尔滨150000

出　　处：《中国医学装备》2020年第9期206-212,共7页China Medical Equipment

基　　金：国家自然科学基金面上项目(81173599)“基于DPC4的蝙蝠葛酚性碱抗胰腺癌分子调控机制研究”;黑龙江省博士后基金(2015M581496)“基于DPC4RANi探讨蝙蝠葛碱联合艾灸对人胰腺癌细胞株Bx PC-3裸鼠移植瘤的影响及机理研究”;黑龙江省普通本科高等学校青年创新人才培养计划(UPYSCT-2018231)“桦褐孔菌醇对BGC-823胃癌细胞RAS-MAPK信号通路相关基因表达调控的影响”。

摘　　要：目的:研究桦褐孔菌(IO)醇提物(IO醇提物)对胃癌细胞BGC-823侵袭转移的影响,以及磷脂酰肌醇3-激酶(PI3K/AKT)通路在其中发挥的作用。方法:药物干预后用四甲基偶氮唑(MTT)比色法检测高剂量组、中剂量组、低剂量组和阴性对照组细胞的增殖情况,划痕实验比较各组细胞的迁移能力,通过细胞迁移(Transwell)实验比较胃癌细胞BGC-823的侵袭能力,采用流式细胞术检测细胞的凋亡及细胞周期情况,逆转录-聚合酶链反应(RT-PCR)和蛋白印迹方法(Westernblot)检测PI3K/AKT通路相关基因、蛋白的表达。观察关键蛋白、基因与IO醇提物抗胃癌的关系。结果:MTT法显示IO醇提物对人胃癌细胞系BGC-823细胞具有抑制作用,高剂量组抑制率为(88.67±2.82)%;流式细胞术观察细胞表明IO醇提物对胃癌BGC-823细胞的凋亡和细胞周期可产生影响,大量胃癌细胞停留于G0/G1期;细胞划痕和Transwell小室表明其对胃癌BGC-823细胞的迁移和侵袭具有抑制作用,细胞迁移和侵袭能力随IO醇提物浓度增高而减弱,侵袭力高剂量组为(40±2.2)%,阴性对照组为(80±3.5)%,其差异有统计学意义(F=166.164,P<0.05);RT-PCR显示抑癌基因PTEN增加,胃癌发生发展相关的mTORmRNA减少,高剂量组分别为(0.91±0.06)bp和(1.65±0.08)bp,阴性对照组分别为(0.64±0.08)bp和(2.38±0.13)bp,其差异有统计学意义(F=18.859,F=55.021;P<0.05);Westernblot法可见PI3K和p-AKT蛋白减少,高剂量组分别为(1.96±0.05)KD和(1.68±0.32)KD,而阴性对照组分别为(6.06±0.35)KD和(6.35±0.73)KD,IO醇提物与阴性对照组PI3K比较差异有统计学意义(F=799.507,F=59.018;P<0.05)。结论:IO醇提物对胃癌BGC-823细胞的迁移侵袭能力具有抑制作用,其发生机制可能与IO醇提物对PI3K/AKT通路有关。Objective: To study the effect of alcohol extract of Inonotus obliquus(IO) on invasion and metastasis of gastric cancer cell BGC-823 and the role of PI3 K/AKT pathway in the process. Methods: Methyl thiazolyl tetrazolium(MTT) method was used to detect the proliferation of cells in each group after drug intervention, and scratch test was used to compare the migration ability of cells in each group. Transwell test was used to compare the invasion ability of gastric cancer cell BGC-823, and flow cytometry(FCM) was used to detect the apoptosis and cell cycle of BGC-823, and reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were used to detect the related gene and the protein expression of PI3 K/AKT pathway. And the relations between key protein and IO alcohol extract, and between key gene and IO alcohol extract that could resist gastric carcinoma were further observed. Results: The results of MMT method indicated IO alcohol extract had inhibiting effect on BGC0823 cell, and the inhibiting rate of high dose group was(88.67±2.82)%. And the results of FCM indicated that IO alcohol extract could produce effects on the apoptosis and cell cycle of gastric carcinoma cell BGC-823, and most of gastric carcinoma cell remained in G0/G1 phase. The results of scratch test and transwell test indicated that IO alcohol extract had inhibiting effect on the migration and invasion of gastric carcinoma cell BGC-823, and the abilities of cell migration and invasion gradually receded with the enhancing of the concentration of IO alcohol extract, and the differences of invasion ability between high-dose group(40±2.2)% and negative control group(80±3.5)% was significant(F=166.164, P<0.05). The results of RT-PCR indicated that the mTOR mRNA that was related with the happen of gastric carcinoma would decrease when tumor suppressor gene PTEN increased, and the tumor suppressor gene PTEN and mTOR mRNA were(0.91±0.06) bp and(1.65±0.08) bp in high dose group, respectively. And they were(0.64±0.08) bp and(2.38±0.13) b

关 键 词：胃癌 桦褐孔菌(IO) 侵袭 迁移 磷脂酰肌醇3-激酶(PI3K/AKT) 

分 类 号：R735.2[医药卫生—肿瘤]