莲房原花青素调控RAC1/PI3K/AkT信号途径抑制结肠癌的增殖、迁移与侵袭  

Lotus seed procyanidin inhibits the proliferation,migration and invasion of colon cancer by inhibiting Rac1/PI3K/Akt signaling pathway

作　　者：张毅 马丹丹 龚齐 黄华 曹定 尚作宏 张翌 ZHANG Yi;MA Dan-dan;GONG Qi;HUANG Hua;CAO Ding;SHANG Zuo-hong;ZHANG Yi(Department of Pharmacy,Wuhan First Hospital,Wuhan 430070,Hubei,China;Department of General Surgery,General Hospital of the Central Theater Command of the People's Liberation Army,Wuhan 430070,Hubei,China)

机构地区：[1]武汉市第一医院药学部,武汉430070 [2]解放军中部战区总医院普通外科,武汉430070

出　　处：《医学研究生学报》2021年第6期580-585,共6页Journal of Medical Postgraduates

基　　金：湖北省卫生健康委员会联合基金项目(WJ 2019H109);湖北省自然科学基金项目(2018CFC803)。

摘　　要：目的莲房原花青素(LSPCs)在结肠癌中的作用目前仍不清楚。文章研究拟探讨LSPCs在结肠癌中的抗肿瘤作用。方法先将不同浓度的LSPCs干预HCT116结肠癌细胞系,实验分为对照组(不予处理)和LSPCs低、中、高浓度组(分别给予10、20、40μmol/L浓度的LSPCs干预24 h),再行CCK-8、Transwell及Western blot检测细胞增殖、转移及侵袭能力。再将HCT116结肠癌细胞系中敲低RAC-1,实验又分为HCT116细胞对照组(不予处理)和si-RAC组(敲低RAC1),24 h后检测细胞的活力与凋亡变化。结果CCK8实验表明莲房原花青素干预后细胞的增殖水平显著下降,并且时间越长,增殖能力越弱(P<0.05);流式凋亡实验结果表明,莲房原花青素干预后细胞的凋亡水平较对照组显著增加(P<0.05)。RT-PCR实验检测表明,与对照组比较,莲房原花青素干预可以显著下调抑凋亡蛋白Bcl-2,并且显著上调促凋亡蛋白Bax表达水平,同时增加Caspase-3表达(P<0.05)。细胞迁移实验表明LSPCs低、中、高浓度组结肠癌细胞侵袭细胞数量(68.45±11.47、52.01±9.43、42.41±8.67)显著低于对照组(97.33±14.32),细胞迁移数量(72.31±10.32、58.04±9.88、49.29±10.05)亦低于对照组(92.67±15.06),差异有统计学意义(P<0.05)。CCK8实验结果表明,与对照组比较,敲低RAC1后细胞的增殖能力显著下调(P<0.05),且同时流式细胞凋亡检测结果证实敲低RAC1后细胞凋亡显著增加,差异有统计学意义(P<0.05)。Transwell实验表明,与HCT116细胞对照组比较,si-RAC组明显下调了HCT116细胞的迁移与侵袭(P<0.05)。Western blot实验结果显示,与HCT116细胞对照组比较,si-RAC组PI3K/AKT的活化显著降低(P<0.05)。结论LSPCs在结肠癌中具有显著的抑制肿瘤恶性生物学行为的作用,促进了结肠癌细胞的凋亡,抑制其增殖、迁移与侵袭,其具体机制可能是通过抑制RAC1/PI3K/AkT信号传导途径实现。Objective Lotus seed procyanidin(LSPC)is one of the main active components isolated from the mature receptacle of Nymphaeaceae,but its role in colon cancer is still unclear.Therefore,this study aims to explore the anti-tumor effect of LSPC in colon cancer.Methods Different concentrations of LSPCs(10,20,40μmol/L)were used to intervene HCT116 colon cancer cell line.Cell apoptosis was detected by flow cytometry,and cell proliferation was detected by CCK8.Transwell assay was used to detect the changes of migration and invasion after LSPCs(10,20,40μmol/L)intervention.Western bolt was used to detect the expression of apoptosis related proteins and Rac-1/PI3K/Akt signaling pathway proteins.Rac-1 was knocked down in HCT116 colon cancer cell line,and cell viability and apoptosis were detected.Results Compared with the control group,LSPCs significantly inhibited cell proliferation and promoted the apoptosis with a dosage effect(P<0.05).Compared with the control group,LSPCs also inhibited the invasion and migration of colon cancer cells with a dosage effect(P<0.05).The expression of Rac-1,p-pi3k and p-Akt were down-regulated after LSPCs intervention(P<0.05).At the same time,knockdown of Rac-1 inhibited the proliferation,migration and invasion of colon cancer,and promoted its apoptosis(P<0.05).Conclusion LSPCs can significantly inhibit the malignant biological behavior of colon cancer,promote the apoptosis of colon cancer cells,inhibit the proliferation,migration and invasion via the inhibition of Rac1/PI3K/Akt signaling pathway.

关 键 词：结肠癌 莲房原花青素 PI3K AKT RAC1 增殖 

分 类 号：R735.35[医药卫生—肿瘤]