摘要
目的研究表明局部麻醉药物普鲁卡因(PCA)可抑制肺癌、肝癌、胃癌、乳腺癌等多种肿瘤细胞的增长,具有抑癌作用。但其对肾癌细胞的影响研究较少。文章旨在研究PCA对肾癌786-O细胞增殖、凋亡的影响及潜在机制。方法实验分为对照组(不加PCA)、PCA组(5 mmol/L PCA)、si-NC组[转染性别决定区Y-box蛋白2(Sex-determining region Y-box protein,SOX2)干扰对照质粒]、si-SOX2组(转染SOX2干扰质粒)、PCA-M+pcDNA3.1组(转染SOX2过表达对照质粒+5 mmol/L PCA)和PCA-M+pcDNA3.1-SOX2组(转染SOX2过表达质粒+5 mmol/L PCA)。MTT法和流式细胞术分别测定细胞增殖和凋亡情况;Western blot测定SOX2、Cyclin D1、Bax和Bcl-2的表达。结果与si-NC组相比,si-SOX2组786-O细胞中SOX2、Cyclin D1和Bcl-2表达量显著降低(P<0.05),Bax表达升高(P<0.05),细胞A 490值在24、48和72 h均显著降低(P<0.05),细胞凋亡率显著升高(P<0.05)与对照组相比,PCA组786-O细胞中SOX2和CyclinD1表达量显著降低(P<0.05),p21蛋白表达量显著升高(P<0.05),细胞A 490值在24、48和72 h均显著降低(P<0.05)。与PCA-M+pcDNA3.1组SOX2(0.24±0.02)和CyclinD1(0.31±0.03)含量相比,PCA-M+pcDNA3.1-SOX2组(0.51±0.05,0.60±0.06)升高(P<0.05),PCA+pcDNA3.1-SOX2组细胞A 490值在24、48和72 h(0.48±0.05、0.68±0.07、0.98±0.10)较PCA+pcDNA3.1组(0.27±0.03、0.40±0.04、0.52±0.05)均显著升高(P<0.05)。与对照组相比,PCA组786-O细胞中Bcl-2表达量显著降低(P<0.05),Bax蛋白表达量显著升高(P<0.05),细胞凋亡率显著升高(P<0.05);与PCA+pcDNA3.1组Bcl-2(0.38±0.04)、Bax(0.62±0.06)表达量、细胞凋亡率[(20.05±2.15)%]相比,PCA+pcDNA3.1-SOX2组786-O细胞中Bcl-2(0.78±0.08)显著升高(P<0.05),Bax(12.29±1.36)表达量显著降低(P<0.05),细胞凋亡率[(12.29±1.36)%]显著降低(P<0.05)。结论PCA通过抑制SOX2表达抑制肾癌786-O细胞增殖并促进细胞凋亡。PCA对肾癌具有潜在抑制作用。
Objective Studies have shown that the local anesthetic drug procaine(procaine,PCA)can inhibit the growth of lung cancer,liver cancer,gastric cancer,breast cancer and other tumor cells,and has cancer suppressing effect.However,its effect on kidney cancer cells has not been studied.The purpose of this experiment was to study the effect and potential mechanism of PCA on the proliferation and apoptosis of renal cancer 786-O cells.Methods The experiment was divided into control(Con)group(without PCA),PCA group(5 mmol/L PCA),si-NC group(transfected with SOX2 interference control plasmid),si-SOX2 group(transfected with SOX2 interference plasmid),PCA-M+pcDNA3.1 group(transfected with SOX2 overexpression control plasmid+5 mmol/L PCA)and PCA-M+pcDNA3.1-SOX2 group(transfected with SOX2 overexpression plasmid+5 mmol/L PCA).MTT method and flow cytometry were used to measure cell proliferation and apoptosis respectively;Western blot was used to determine the expression of sex-determining region Y-box protein 2(SOX2),Cyclin D1,Bax and Bcl-2.Results Compared with si-NC group,the expression of SOX2,Cyclin D1 and Bcl-2 in 786-O cells from si-SOX2 group was significantly decreased(P<0.05),expression of Bax was increased(P<0.05),A490 value of cells was significantly decreased at 24,48 and 72 h(P<0.05),and the apoptosis rate was significantly increased(P<0.05).In comparison with the control group,there was a significant reduction(P<0.05)in SOX2 and Cyclin D1 expression of 786-O cells in PCA group,and a significant increase in the expression of p21 protein(P<0.05).A490 value of cells reduced significantly at 24,48 and 72 h(P<0.05).The content of SOX2(0.51±0.05)and cyclin D1(0.60±0.06)in PCA-M+pcDNA3.1-SOX2 group was higher(P<0.05)than that in PCA-M+pcDNA3.1 group(0.24±0.02,0.31±0.03).A490 value of cells in PCA+pcDNA3.1-SOX2 group at 24,48 and 72 h(0.48±0.05,0.68±0.07,0.98±0.10)was significantly higher(P<0.05)than that in PCA+pcDNA3.1 group(0.27±0.03,0.40±0.04,0.52±0.05).Compared with the control group,in PCA group,the expres
作者
余晖
宋辉琼
程江霞
彭晓红
YU Hui;SONG Hui-qiong;CHENG Jiang-xia;PENG Xiao-hong(Department of Anesthesiology,Fourth Hospital of Wuhan/Pu'ai Hospital of Tongji Medical College Affiliated to Huazhong University of Science and Technology,Wuhan 430033,Hubei,China)
出处
《医学研究生学报》
CAS
北大核心
2020年第11期1152-1156,共5页
Journal of Medical Postgraduates
