结直肠癌细胞在三维与二维培养环境下增殖机制研究  

Study on the proliferation mechanism of colorectal cancer cells in three-dimensio and two-dimensio culture environment

作　　者：胡艳艳 秦宝丽[1] 周璐[2] 邱月 椰慧楠[1] 张欣慰 HU Yan-yan;QIN Bao-li;ZHOU Lu;QIU Yue;YE Hui-nan;ZHANG Xin-wei(The First Department of Digestive Diseases,China Medical University Cancer Hospital Liaoning Cancer Hospital,Shenyang 110042,China;Department of Thoracic and Esophageal Radiotherapy,China Medical University Cancer Hospital Liaoning Cancer Hospital,Shenyang 110042,China;Integrated Chinese and Western Medicine,China Medical University Cancer Hospital Liaoning Cancer Hospital,Shenyang 110042,China)

机构地区：[1]中国医科大学肿瘤医院·辽宁省肿瘤医院消化内科一病区,辽宁沈阳110042 [2]中国医科大学肿瘤医院·辽宁省肿瘤医院胸部·食管放疗病区,辽宁沈阳110042 [3]中国医科大学肿瘤医院·辽宁省肿瘤医院中西医结合科,辽宁沈阳110042

出　　处：《临床军医杂志》2024年第1期42-45,共4页Clinical Journal of Medical Officers

基　　金：辽宁省博士科研启动基金(2020-BS-042)。

摘　　要：目的 探讨结直肠癌细胞在三维(3D)与二维(2D)培养环境下增殖能力的变化及机制。方法 SW480结直肠癌细胞购自上海细胞库。将SW480结直肠癌细胞分别进行壳聚糖-胶原水凝胶3D培养与细胞培养皿2D培养。光学显微镜观察细胞形态;采用细胞计数试剂盒(CCK-8)检测细胞增殖情况;Western blot法检测3D与2D培养环境OTUB2与PCNA蛋白表达;比较转染前后SW480细胞OTUB2、PCNA基因和蛋白表达、细胞增殖速率。结果 SW480结肠癌细胞在2D环境培养24 h后可圆形或卵圆形细胞贴壁,7 d后已贴壁的细胞慢慢伸展,呈短杆状;SW480结肠癌细胞在壳聚糖-胶原水凝胶3D培养24 h后呈3D球状生长,培养7 d后可见细胞数量增多,细胞球变大。3D培养环境SW480细胞增殖速率低于2D培养环境,差异有统计学意义(P<0.05)。Western blot法检测结果显示,与2D培养比较,3D培养环境培养后结直肠癌细胞系中OTUB2、PCNA蛋白表达均低于2D培养环境,差异均有统计学意义(P<0.05)。转染后SW480细胞OTUB2、PCNA基因和蛋白表达、细胞增殖速率均高于转染前,差异均有统计学意义(P<0.05)。结论 壳聚糖-胶原水凝胶3D培养条件下结直肠癌细胞增殖速率减慢,其机制与调控OTUB2、PCNA表达有关。Objective To investigate the change and mechanism of proliferation ability of colorectal cancer cells in three-dimensio(3D) and two-dimensio(2D) culture environment.Methods SW480 colorectal cancer cells were purchased from Shanghai Cell Bank.SW480 colorectal cancer cells were cultured by chitosan-collagen hydrogel 3D and cell culture dish 2D respectively.The cell morphology was observed by optical microscope.Cell proliferation was detected with cell counting kit-8(CCK-8)cell counting kit.The expressions of OTUB2 and PCNA in 3D and 2D culture were detected by Western blot.The expression of OTUB2,PCNA gene and protein and cell proliferation rate of SW480 cells before and after transfection were compared.Results SW480 colon cancer cells could stick to the wall after 24 hours in 2D environment,and the attached cells slowly spread out and showed short rod shape after 7 days.SW480 colon cancer cells grew in 3D spheres after 3D culture of chitosan-collagen hydrogel for 24 hours,and the number of cells increased and the cell spheres became larger after 7 days of culture.The proliferation rate of SW480 cells in 3D culture environment was lower than that in 2D culture environment,and the difference was statistically significant(P<0.05).Western blot analysis showed that compared with 2D culture,the expression of OTUB2 and PCNA protein in colorectal cancer cell lines cultured in 3D culture environment was lower than that in 2D culture environment,and the differences were statistically significant(P<0.05).The expression of OTUB2,PCNA gene and protein and cell proliferation rate of SW480 cells after transfection were higher than those before transfection,with statistical significance(P<0.05).Conclusion The proliferation rate of colorectal cancer cells slowed down under the condition of chitosan-collagen hydrogel 3D culture,and the mechanism is related to the regulation of OTUB2 and PCNA expression.

关 键 词：三维培养 二维培养 蛋白表达 结直肠癌 细胞增殖 

分 类 号：R735.34[医药卫生—肿瘤]