摘要
目的 评估抗体偶联药物维迪西妥单抗(RC48)对不同HER-2蛋白表达水平胃癌细胞的体外抑制效应。方法 使用实时荧光定量PCR(qRT-PCR)、Western blotting技术结合细胞免疫荧光共聚焦显微镜,检测胃癌细胞系(NCI-N87、MKN45、MKN7、HGC27、MGC803)HER-2表达情况及细胞定位。随后对上述不同HER-2表达程度的细胞系分别经不同浓度RC48处理,CCK-8技术检测杀伤效应并计算半数抑制浓度(IC50),采用qRT-PCR和Western blotting技术检测胃癌细胞经RC48处理后,不同胃癌细胞株的HER-2 mRNA及蛋白质表达情况。结果 所检测的胃癌细胞系中,NCI-N87细胞的HER-2表达量最高,其余细胞系均呈HER-2低表达,荧光共聚焦显示HER-2蛋白主要为细胞膜定位。RC48在体外可明显抑制HER-2强阳性胃癌细胞(NCI-N87)增殖,48 h及72 h的IC50分别为0.32μg/ml及2.359e-010μg/ml,该效应呈现明显的剂量依赖性和时间依赖性;而在HER-2低表达胃癌细胞(MKN45、MKN7、HGC27、MGC803)中,RC48同样显示了一定的体外抑制肿瘤细胞增殖的效应,48 h的IC50分别为331.90、24.20、35.32及9.449μg/ml。不论基线时的HER-2蛋白表达高低,经RC48处理后胃癌细胞的HER-2蛋白表达量均呈明显下降。结论 RC48对体外培养的HER-2不同表达胃癌细胞均显示了抑制增殖的效应,呈时间依赖性和剂量依赖性。RC48作用后可抑制胃癌细胞表面的HER-2蛋白表达,且该效应并不完全依赖于胃癌细胞基线HER-2蛋白表达量的高低。
Objective To evaluate the antitumor activity and the potential beneficiaries of HER-2-antibody drug conjugate(Disitamab Vedotin, RC48) in gastric cancer cells with different HER-2 expression levels. Methods Human NCI-N87, MKN45, MKN7, HGC27, MGC803 gastric cancer cell lines were detected by real time fluorescent quatitative PCR(qRT-PCR), Western blotting combined with confocal cell immunofluorescence to expose their expression and cellular localization of HER-2 in gastric cancer. Subsequently, the above cell lines were treated with different concentrations of RC48 to detect lethal effect and evaluate half inhibitory concentration(IC50) by using cell counting kit(CCK)-8 assay in vitro gastric cancer models. In addition, qRT-PCR and Western blotting techniques were performed to detect the HER-2 mRNA and protein expression level of different gastric cancer cell lines after exposed to RC48. Results Among the gastric cancer cell lines tested, NCI-N87 cells had the highest expression of HER-2, while the rest of the cell lines showed weak HER-2 expression. The immunofluorescence confocal showed that HER-2 was mainly located in the cell membrane. RC48 had a dose and time-dependent inhibitory effect on HER-2 strongly positive gastric cancer cells(NCI-N87), with IC50of 48 h and 72 h showed 0.32 μg/ml and 2.359e-010μg/ml, respectively. At the same time, it also showed certain antitumor effect in HER-2 low expression cell models like MKN45, MKN7, HGC27, MGC803, with IC50of 48 h showed 331.90, 24.20, 35.32 and 9.449 μg/ml, respectively. Regardless of the level of HER-2 protein expression at baseline, the expression of HER-2 in gastric cancer cells after RC48 treatment was significantly decreased. Conclusion RC48 can inhibit the proliferation of different gastric cells with different expression levels of HER-2 in a certain time and dose-manner. At the same time, RC48 can also inhibit the expression of HER-2 protein in gastric cancer cells, which efficacy does not completely depend on the level of baseline HER-2 expression.
作者
金洋冰
蔡劬
计骏
施敏
张俊
JIN Yangbing;CAI Qu;JI Jun;SHI Min;ZHANG Jun(Department of Oncology,Ruijin Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China)
出处
《临床肿瘤学杂志》
CAS
2023年第1期1-7,共7页
Chinese Clinical Oncology
基金
国家自然科学基金资助项目(81972707,82273126)
上海市卫生健康委协同创新集群计划(2020CXJQ03)。
