摘要
目的探讨奥利司他对人胆囊癌细胞活性的影响。方法采用实验研究方法。通过体外培养人胆囊癌NOZ、GBC‑SD、SGC‑996细胞,筛选出高表达FASN的人胆囊癌NOZ细胞采用细胞增殖、细胞克隆形成和蛋白检测实验,分析奥利司他对人胆囊癌细胞活性的影响。人胆囊癌NOZ细胞分组:加入培养基设为NOZ细胞对照组,加入培养基+10μmol/L奥利司他设为奥利司他低浓度组;加入培养基+100μmol/L奥利司他,设为奥利司他高浓度组。观察指标:(1)FASN蛋白在人胆囊癌细胞中的表达情况。(2)奥司利他对人胆囊癌NOZ细胞增殖的影响。(3)奥司利他对人胆囊癌NOZ细胞凋亡的影响。正态分布的计量资料以x±s表示,组间比较采用AVONA检验,两两比较采用最小显著差异法。结果(1)FASN蛋白在人胆囊癌细胞中的表达情况。Western blot检测结果显示:FASN蛋白在人胆囊癌NOZ、GBC‑SD、SGC‑996细胞中的相对表达量分别为0.57±0.06、0.12±0.04、0.10±0.02,3者比较,差异有统计学意义(F=115.67,P<0.05);NOZ细胞分别与GBC‑SD、SGC‑996细胞比较,差异均有统计学意义(P<0.05)。GBC‑SD细胞和SGC‑996细胞比较,差异无统计学意义(P>0.05)。(2)奥利司他对人胆囊癌NOZ细胞增殖的影响。①细胞增殖毒性实验结果显示:NOZ细胞对照组、奥利司他低浓度组、奥利司他高浓度组的吸光度分别为2.34±0.12、1.57±0.08、1.07±0.13,3组比较,差异有统计学意义(F=205.88,P<0.05)。②细胞克隆形成实验结果显示:NOZ细胞对照组、奥利司他低浓度组、奥利司他高浓度组的细胞克隆形成数目分别为(257±23)个、(153±11)个、(83±11)个,3组比较,差异有统计学意义(F=92.64,P<0.05)。③Western blot实验结果显示:NOZ细胞对照组、奥利司他低浓度组、奥利司他高浓度组中Cyclin‑D1蛋白的相对表达量分别为2.31±0.10、1.52±0.05、1.23±0.11,3组比较,差异有统计学意义(F=120.73,P<0.05)。NOZ细胞对照组、奥利司他低浓度组、奥利司他高浓度组中CDK‑4蛋白的相对表达量分别为1.58±0.04、1.21±0.02、1.19±0.04,3组比较,差异有统计学意义(F=110.45,P<0.05)。(3)奥利司他对人胆囊癌NOZ细胞凋亡的影响。Western blot实验结果显示:NOZ细胞对照组、奥利司他低浓度组、奥利司他高浓度组中Bcl‑2蛋白的相对表达量分别为1.07±0.03、0.36±0.03、0.15±0.02,3组比较,差异有统计学意义(F=1242.93,P<0.05)。NOZ细胞对照组、奥利司他低浓度组、奥利司他高浓度组中Bax蛋白的相对表达量分别为0.51±0.03、0.38±0.05、1.38±0.04,3组比较,差异有统计学意义(F=583.51,P<0.05)。结论奥利司他可以抑制人胆囊癌NOZ细胞的生长,促进其凋亡。
Objective To investigate the effects of orlistat on the viability of human gallbladder cancer(GBC)cells.Methods The experimental study was conducted.The human GBC NOZ cells with high expression of FSAN was screened out through in vitro cultivating human GBC-SD,SGC-996 and NOZ cells.The cell proliferation assay,clone formation assay and protein detection experiment were used to analysis of the effects of orlistat on the viability of human GBC cells.Cell grouping:NOZ cells cultured with medium were set as the control group,cultured with medium+10μmol/L orlistat were set as the low-dose orlistat group,cultured with medium+100μmol/L orlistat were set as the high-dose orlistat group,respectively.Observation indicators:(1)expression of FASN protein in human GBC cells;(2)effects of orlistat on the proliferation of human GBC NOZ cells;(3)effects of orlistat on apoptosis of human GBC NOZ cells.Measurement data with normal distribution were represented as Mean±SD,the ANOVA test was used for comparison between groups and the least significant difference method was used for pairwise comparison.Results(1)Expression of FASN protein in human GBC cells.Results of western blot showed that the relative expression of FASN protein in human GBC NOZ,GBC-SD and SGC-996 cells was 0.57±0.06,0.12±0.04 and 0.10±0.02,respectively,showing a significant difference among them(F=115.67,P<0.05).There were significant differences between the NOZ cells and the GBC-SD or the SGC-996 cells(P<0.05),and there was no significant difference between the GBC-SD cells and the SGC-996 cells(P>0.05).(2)Effects of orlistat on the proliferation of human GBC NOZ cells.①Results of cell proliferation assay showed that the absorbance value of NOZ cells was 2.34±0.12,1.57±0.08 and 1.07±0.13 in the control group,lowdose orlistat group and high-dose orlistat group,respectively,showing a significant difference among them(F=205.88,P<0.05).②Results of clone formation assay showed that the number of NOZ cells clones was 257±23,153±11 and 83±11 in the control group,low-dose orlistat group and high-dose orlistat group,respectively,showing a significant difference among them(F=92.64,P<0.05).③Results of western blot showed that the relative expression of Cyclin-D1 protein of NOZ cells was 2.31±0.10,1.52±0.05 and 1.23±0.11 in the control group,low-dose orlistat group and high-dose orlistat group,respectively,showing a significant difference among them(F=120.73,P<0.05).The relative expression of CDK-4 protein of NOZ cells was 1.58±0.04,1.21±0.02 and 1.19±0.04 in the control group,low-dose orlistat group and high-dose orlistat group,respectively,showing a significant difference among them(F=110.45,P<0.05).(3)Effects of orlistat on apoptosis of human GBC NOZ cells.Results of western blot showed that the relative expression of Bcl-2 protein of NOZ cells was 1.07±0.03,0.36±0.03 and 0.15±0.02 in the control group,low-dose orlistat group and high-dose orlistat group,respectively,showing a significant difference among them(F=1242.93,P<0.05).The relative expression of Bax protein of NOZ cells was 0.51±0.03,0.38±0.05 and 1.38±0.04 in the control group,low-dose orlistat group and high-dose orlistat group,respectively,showing a significant difference among them(F=583.51,P<0.05).Conclusion Orlistat can inhibit the growth of human GBC NOZ cells and promote their apoptosis.
作者
程海鸿
孙玉鑫
于小鹏
王寿华
丁俊
周迪
张晓宇
王健东
施伟斌
马飞
Cheng Haihong;Sun Yuxin;Yu Xiaopeng;Wang Shouhua;Ding Jun;Zhou Di;Zhang Xiaoyu;Wang Jiandong;Shi Weibin;Ma Fei(Department of Oncology,Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine,Shanghai 200092,China;Department of General Surgery,Xinhua Hospital Affiliated to Shanghai Jiaotong University School of Medicine,Shanghai 200092,China;Department of Biliary and Pancreatic Surgery,Shanghai Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China;Department of Hepatobiliary Surgery,Tenth People's Hospital of Tongji University,Shanghai 200072,China)
出处
《中华消化外科杂志》
CAS
CSCD
北大核心
2023年第5期636-641,共6页
Chinese Journal of Digestive Surgery
基金
上海市卫健委临床研究专项面上项目(202040447)。
关键词
胆囊肿瘤
奥利司他
细胞增殖
细胞凋亡
细胞活性
周期蛋白
凋亡相关蛋白
Gallbladder neoplasms
Orlistat
Cell proliferation
Cell apoptosis
Cell viability
Cyclins
Apoptosis-related proteins
作者简介
通信作者:马飞,Email:mafei@xinhuamed.com.cn。
