中国药理学通报2024，Vol.40Issue(4)：652-661,10.DOI:10.12360/CPB202309050
血根碱对患者源性结直肠癌类器官生长的影响
Effect of sanguinarine on organoids growth in patients with colorectal cancer
摘要
Abstract
Aim To investigate the effect of sanguina-rine(SNG)on organoid growth in patients with color-ectal cancer and its molecular mechanism.Methods Tumor tissues were collected from resected colorectal carcinoma and the tumor organoids were cultured in vitro.The dynamic changes of organoid formation in colorectal carcinoma were observed under light micro-scope.The intercellular structure of colorectal carcino-ma was observed by HE staining.The expression of tumor markers in colorectal carcinoma was observed by immunohistochemical staining.SNG at different con-centrations(0.1,0.3,1,3,9,27 μmol·L-1)was used to process organoids.The cell activity was meas-ured by CCK-8 method and the IC50 was calculated.The effect of SNG on the expression of organoid pro-teins in colorectal cancer was detected by Western blot.ATP content was used to detect the synergistic effect of SNG combined with chemotherapeutic drugs on organoids.Results Three-dimensional cell clus-ters with cystoid and parenchymatous structure were formed at 48-72 h,suggesting that organoids were successfully constructed.The expression of Ki67,CK20,CK7 and Ep-CAM in organoids was confirmed by immunohistochemistry.The results of CCK-8 showed that compared with the control group,the num-ber of organoids and the activity of cells in the SNG group were decreased.Western blot showed that the expression of p-AMPK was significantly increased,while the expression of mTOR and p-mTOR was signifi-cantly decreased(P＜0.05)in a concentration-de-pendent manner.The expression of autophagy-related proteins p62 and Beclin1 also significantly changed(P＜0.05).The results of ATP content assay showed that the inhibition of proliferation of colorectal cancer cells by SNG combined with clinical chemotherapeutic drugs was significantly higher than that of clinical che-motherapeutic drugs alone(P＜0.05).Conclusions SNG can inhibit the growth of colorectal cancer or-ganoids and induce autophagy.SNG,together with the oxaliplatin of clinical chemotherapy drugs,can inhibit the proliferation of tumor cells,and the mechanism may be related to the regulation of AMPK/mTOR sig-naling pathway.关键词
血根碱/结直肠癌/类器官/AMPK/自噬/临床化疗药Key words
sanguinarine/colorectal cancer/or-ganoid/AMPK/autophagy/clinical chemotherapy drugs分类
医药卫生引用本文复制引用
徐志东,郑海伦,周军伟,倪超,柯希权,马振增,邓晓晶,赵睿,顾林,任志..血根碱对患者源性结直肠癌类器官生长的影响[J].中国药理学通报,2024,40(4):652-661,10.基金项目
安徽省高等学校科学研究项目(自然科学类)重大项目(No 2022AH040216) （自然科学类）
安徽高校自然科学研究项目(No KJ2021A0813) （No KJ2021A0813）
蚌埠医学院512人才培育计划项目(No by51201216) （No by51201216）
蚌埠医学院研究生科研创新计划项目(No Byycx22027) （No Byycx22027）
