摘要
目的研究桦褐孔菌醇提物(IO醇提物)对胃癌BGC-823裸鼠移植瘤的抑制作用,推测其作用机制与PI3K/AKT通路相关性。方法48只裸鼠使用异位移植瘤造模法建立胃癌模型,造模后按照随机数字法分为IO醇提物高剂量组(3 mg/g)、中剂量组(1 mg/g)、低剂量组(0.75 mg/g),阴性对照组(羧甲基纤维素钠灌胃),模型组(不灌药物),阳性对照组(卡培他滨灌胃,0.5 mg/g),每组各8只。灌胃14 d,处死小鼠,剥离肿瘤组织,称重计算抑瘤率;光镜和电镜下观察药物干预后肿瘤组织形态变化;免疫组织化学法检测相关蛋白4E-BP1、p27kip1、Bcl-2、Bax、caspase-3的表达;逆转录-聚合酶链式反应法(RT-PCR)检测磷酸酶基因(PTEN)、磷脂酰肌醇-3-羟激酶(PI3K)、蛋白激酶(AKT)、雷帕霉素靶蛋白(mTOR)表达;蛋白免疫印迹法(Western blot)测定PI3K、p-AKT蛋白表达。结果与阴性对照组、模型组比较,各IO醇提物组肿瘤重量降低(P<0.05),生长受到抑制(P<0.05);各IO醇提物组的肿瘤坏死、凋亡、细胞器破坏明显;与阴性对照组、模型组比较,各IO醇提物组Bcl-2细胞平均光密度值降低,4E-BP1、p27kip1、Bax、caspase-3细胞平均光密度值升高(P<0.05);RT-PCR检测结果示,PTEN在各IO醇提物组表达均高于阴性对照组、模型组,低于阳性对照组(P<0.05),mTOR、PI3K和AKT表达低于阴性对照组、模型组,高于阳性对照组(P<0.05)。Western blot检测结果示,各IO醇提物组PI3K、p-AKT蛋白表达低于阴性对照组、模型组,高于阳性对照组(P<0.05)。结论IO醇提物具有较为明显的胃癌抑制作用,其机制可能与PI3K/AKT通路及其上下游因子有关。
Objective To investigate the inhibitory effect of Alcohol extract of Inonotus Obliquus(Alcohol extract of IO)on transplanted tumor of gastric cancer BGC-823 in nude mice,and speculate the relationship between the mechanism and PI3K/AKT pathway.Methods Forty-eight nude mice were used to establish gastric cancer models by heterotopic transplantation.After modeling,they were randomly divided into Alcohol extract of IO high-dose group(3 mg/g),middle-dose group(1 mg/g)and low-dose group(0.75 mg/g),negative control group(hydroxymethyl cellulose sodium lavage),model group(no drug),positive control group(Capecitabine lavage,0.5 mg/g).according to the random number table method,with 8 mice in each group.Mice were sacrificed after 14 days of intragastric administration,tumor tissues were stripped and tumor inhibition rate was calculated by weighing.The morphological changes of tumor tissues after drug intervention were observed under light microscope and electron microscope.The expressions of 4E-BP1,p27kip1,Bcl-2,Bax and caspase-3 were detected by immunohistochemistry.The expressions of Phosphatase gene(PTEN),phosphatidylinositol-3-hydroxykinase(PI3K),protein kinase(AKT),and target of rapamycin(mTOR)were detected by reverse transcription-polymerase chain reaction(RT-PCR).The protein expressions of PI3K and p-AKT were detected by Western blot.Results Compared with the negative control group and model group,the weight of tumor were reduced in Alcohol extract of IO group(P<0.05),the growth in each Alcohol extract of IO groups was inhibited(P<0.05).In the each Alcohol extract of IO groups,tumor necrosis,apoptosis and organelle destruction were more obvious(P<0.05).Compared with the negative control group and model group,the average optical density of Bcl-2 in each Alcohol extract of IO groups was decreased,the average optical density of 4E-Bp1,p27kip1,Bax and caspase-3 was increased(P<0.05).RT-PCR detect results showed that,the expression of PTEN in each Alcohol extract of IO groups was higher than that in the negative control group and model group,and was lower than that in the positive control group(P<0.05);the expressions of mTOR,PI3K and AKT in the each Alcohol extract of IO groups were higher than those in the positive control group,and were lower than those in the negative control group and model group(P<0.05).Western blot detect results showed that,the expressions of PI3K and p-AKT protein in each Alcohol extract of IO groups were lower than those in negative control group and model group,and were higher those that in the positive control group(P<0.05).Conclusion Alcohol extract of IO has obvious inhibitory effect on gastric cancer,and its mechanism may be related to PI3K/AKT pathway and its upstream and downstream factors.
作者
王蔚
周忠光
杨婧
刘旭
田明
乔羽
仲丽丽
WANG Wei;ZHOU Zhongguang;YANG Jing;LIU Xu;TIAN Ming;QIAO Yu;ZHONG Lili(Teaching and Research Department of Pathology,Basic Medical College,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China;Teaching and Research Department of Histoembryology,Basic Medical College,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China;Experiment and Training Center,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China;Teaching and Research Department of Shanghan,Basic Medical College,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China;Department of Pathology,the First Clinical Medical College,Heilongjiang University of Chinese Medicine,Heilongjiang Province,Harbin150040,China)
出处
《中国医药导报》
CAS
2020年第14期8-13,共6页
China Medical Herald
基金
国家自然科学基金资助项目(81173599)
黑龙江省普通本科高等学校青年创新人才培养计划项目(UPYSCT-2018231)
中国博士后科学基金项目(2015M581496)。
