摘要
目的探究长链非编码RNA(LncRNA)COL11A1-208在胃癌(GC)中的表达及对细胞增殖、侵袭和迁移的影响。方法收集49例GC患者的癌组织及癌旁组织标本,体外培养人正常胃黏膜上皮细胞(GES-1)及人GC细胞(AGS、MGC-803、MKN-45、BGC-823、SGC-7901),qRT-PCR检测癌旁组织、癌组织及各GC细胞中COL11A1-208表达情况。对SGC-7901细胞进行转染并将其分为Ctrl组(未进行转染)、pcDNA3.1组(转染pcDNA3.1载体)、pcDNA3.1-COL11A1-208组(转染pcDNA3.1-COL11A1-208)、si-NC组(转染si-NC)、si-COL11A1-208组(转染si-COL11A1-208),qRT-PCR验证转染效果;CCK-8实验、克隆形成实验检测各组SGC-7901细胞增殖;Transwell实验检测各组SGC-7901细胞侵袭;划痕愈合实验检测各组SGC-7901细胞迁移;Western Blot检测各组SGC-7901细胞增殖、侵袭和迁移相关蛋白[Ki67、细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶9(MMP9)、神经型钙黏附蛋白(N-cadherin)]表达。结果与癌旁组织比较,GC癌组织中COL11A1-208表达增加(P<0.05);与GES-1细胞比较,AGS细胞、MGC-803细胞、MKN-45细胞、BGC-823细胞、SGC-7901细胞中COL11A1-208表达均增加(P<0.05),且SGC-7901细胞增加更多;与Ctrl组、pcDNA3.1组、si-NC组比较,pcDNA3.1-COL11A1-208组SGC-7901细胞中COL11A1-208表达增加(P<0.05),SGC-7901细胞增殖率、克隆数、侵袭数、划痕面积愈合率及Ki67、Cyclin D1、MMP9、N-cadherin表达均增加(P<0.05),si-COL11A1-208组SGC-7901细胞中COL11A1-208表达减少(P<0.05),SGC-7901细胞增殖率、克隆数、侵袭数、划痕面积愈合率及Ki67、Cyclin D1、MMP9、N-cadherin表达均减少(P<0.05)。结论COL11A1-208在GC中高表达,其可促进SGC-7901细胞增殖、侵袭和迁移,COL11A1-208可能为GC一个潜在的生物标志物。
Objective To investigate the expression of long non-coding RNA(LncRNA)COL11A1-208 in gastric cancer(GC)and its influences on cell proliferation,invasion and migration.Methods The cancer tissue and adjacent tissue samples of 49 GC patients were collected,and human normal gastric epithelial cells(GES-1)and human GC cells(AGS,MGC-803,MKN-45,BGC-823,SGC-7901)were cultured in vitro,qRT-PCR was used to detect the expression of COL11A1-208 in adjacent tissues,cancer tissues and GC cells.SGC-7901 cells were transfected and grouped into Ctrl group(not transfected),pcDNA3.1 group(transfected with pcDNA3.1 vector),pcDNA3.1-COL11A1-208 group(transfected with pcDNA3.1-COL11A1-208),si-NC group(transfected with si-NC),si-COL11A1-208 group(transfected with si-COL11A1-208),qRT-PCR was used to verify the transfection effect;the proliferation of SGC-7901 cells in each group was detected by CCK-8 experiment and clone formation experiment;Transwell assay was used to detect the invasion of SGC-7901 cells in each group;the migration of SGC-7901 cells in each group was detected by scratch healing assay;Western Blot was performed to detect the expression of SGC-7901 cell proliferation,invasion and migration related proteins[Ki67,Cyclin D1,matrix metalloproteinase 9(MMP9),neural cadherin(N-cadherin)]in each group.Results Compared with adjacent tissues,the expression of COL11A1-208 in GC cancer tissues increased(P<0.05);compared with GES-1 cells,the expression of COL11A1-208 in AGS cells,MGC-803 cells,MKN-45 cells,BGC-823 cells,and SGC-7901 cells all increased(P<0.05),and SGC-7901 cells increased more;compared with Ctrl group,pcDNA3.1 group and si-NC group,the expression of COL11A1-208 in SGC-7901 cells of pcDNA3.1-COL11A1-208 group increased(P<0.05),the SGC-7901 cell proliferation rate,clone number,invasion number,wound healing rate,and the expression of Ki67,Cyclin D1,MMP9,and N-cadherin all increased(P<0.05),the expression of COL11A1-208 decreased in SGC-7901 cells in si-COL11A1-208 group(P<0.05),the SGC-7901 cell proliferation rate,clone number,invasion number,wound healing rate,and expression of Ki67,Cyclin D1,MMP9,and N-cadherin all decreased(P<0.05).Conclusion COL11A1-208 is highly expressed in GC,which can promote the proliferation,invasion and migration of SGC-7901 cells.COL11A1-208 may be a potential biomarker of GC.
作者
陈冲
徐康
童诚
崔建华
张祖芳
CHEN Chong;XU Kang;TONG Cheng;CUI Jianhua;ZHANG Zufang(Department of Gastroenterology,Dongtai People’s Hospital,Dongtai,Jiangsu 224200,China)
出处
《中国实验诊断学》
2023年第11期1320-1325,共6页
Chinese Journal of Laboratory Diagnosis
基金
2020年盐城市医学科技发展计划项目(编号:YK2020108)
