普鲁卡因通过上调lncRNA DGCR5抑制胃癌细胞增殖、迁移和侵袭  

Procaine inhibits the proliferation,migration and invasion of gastric cancer cells through up-regulating lncRNA DGCR5

作　　者：李晶[1] 潘侠[1] 周芳[1] 汪晶 洪佳 Jing Li;Xia Pan;Fang Zhou;Jing Wang;Jia Hong(Department of Anesthesiology,People's Hospital of Wuhan University,Wuhan 430000,China;Obstetrics and Gynecology,People's Hospital of Wuhan University,Wuhan 430000,China)

机构地区：[1]武汉大学人民医院麻醉科,武汉430000 [2]武汉大学人民医院妇产科,武汉430000

出　　处：《中华细胞与干细胞杂志（电子版）》2024年第3期151-158,共8页Chinese Journal of Cell and Stem Cell（Electronic Edition）

基　　金：武汉大学人民医院湖北省重点实验室开放项目(2023KFH008)。

摘　　要：目的探究普鲁卡因在胃癌细胞迁移、侵袭和增殖中的作用及潜在机制。方法胃癌AGS细胞分为普鲁卡因低、中、高剂量组(分别用含3、6、12 mmol/L普鲁卡因的培养基干预AGS细胞24 h)、pcDNA-lncRNA DGCR5组(常规培养基培养转染lncRNA DGCR5过表达载体的AGS细胞24 h)、pcDNA-NC组(常规培养基培养转染空载体的AGS细胞24 h)、高剂量+si-lncRNA DGCR5组(含12 mmol/L普鲁卡因的培养基干预转染lncRNA DGCR5小干扰RNA的AGS细胞24 h)、高剂量+si-NC组(含12 mmol/L普鲁卡因的培养基干预转染乱序无意义阴性序列的AGS细胞24 h)和对照组(AGS细胞常规培养24 h),分别采用MTT、克隆形成、划痕愈合、Transwell和蛋白印迹法检测细胞增殖、迁移和侵袭及细胞中MMP2、MMP9蛋白表达,RT-qPCR法检测lncRNA DGCR5表达。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析和LSD-t检验。结果与对照组比较,普鲁卡因低、中、高剂量组AGS细胞A值、克隆数、划痕愈合率、迁移数和侵袭数及MMP2和MMP9蛋白表达量均降低(P均<0.05),lncRNA DGCR5表达升高(1.69±0.13、2.18±0.17、2.49±0.15比1.00±0.10,P<0.05)。与pcDNA-NC组比较,pcDNA-lncRNA DGCR5组AGS细胞A值、克隆数、划痕愈合率、迁移数和侵袭数及MMP2和MMP9蛋白表达量均降低(P均<0.05)。与高剂量+si-NC组比较,高剂量+si-lncRNA DGCR5组A值、克隆数、划痕愈合率、迁移和侵袭数及MMP2、MMP9蛋白表达增高(P均<0.05)。结论普鲁卡因可能提高lncRNA DGCR5表达来阻碍胃癌AGS细胞的迁移、侵袭和增殖。Objective To explore procaine's role and potential mechanism in the migration,invasion and proliferation of gastric cancer cells.Methods Gastric cancer AGS cells were divided into low,medium,and high dose of procaine groups(cells were cultured in medium containing 3,6,12 mmol/L procaine for 24 h),pcDNA-lncRNA DGCR5 group(AGS cells transfected with lncRNA DGCR5 overexpression vector were cultured in the conventional medium for 24 h),pcDNA-NC group(AGS cells transfected with empty vector were cultured in the conventional medium for 24 h),high-dose+si-lncRNA DGCR5 group(AGS cells transfected with lncRNA DGCR5 small interfering RNA were cultured in medium containing 12 mmol/L procaine for 24 h),high-dose+si-NC group(AGS cells transfected with out-of-order nonsense negative sequence were cultured in medium containing 12 mmol/L procaine for 24 h),and control group(AGS cells were cultured in conventional medium for 24 h).And then,MTT,clone formation,scratch healing,Transwell and Western blot method were used to detect cell proliferation,migration and invasion,and the protein expression of MMP2 and MMP9 in cells,respectively;RT-qPCR was used to detected the expression of lncRNA DGCR5.Independent sample t-test was used for comparison between two groups,and univariate analysis of variance and LSD-t test were used for comparison between multiple groups.Results Compared with the control group,the A value,clone count,scratch healing rate,the number of migration and invasion of AGS cells,and the protein expression of MMP2 and MMP9 in the low,medium and high dose groups of procaine were decreased(P all<0.05),and the expression of lncRNA DGCR5(1.69±0.13,2.18±0.17,2.49±0.15 vs 1.00±0.10)was increased(P<0.05).Compared with the pcDNA-NC group,the A value,clone count,scratch healing rate,the number of migration and invasion of AGS cells,and the protein expression of MMP2 and MMP9 in the pcDNA-lncRNA DGCR5 group were all decreased(P all<0.05).Compared with the high-dose+si-NC group,the A value,clone count,scratch healing rate,the

关 键 词：普鲁卡因 胃癌 lncRNA DGCR5 细胞增殖 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]