[1]李怡君,杨振威,陈晓露,等.Ⅳ型胶原蛋白基因α1对胃癌细胞增殖、转移及凋亡的调控作用与机制[J].新乡医学院学报,2022,39(4):311-317.[doi:10.7683/xxyxyxb.2022.04.003]
 LI Yijun,YANG Zhenwei,CHEN Xiaolu,et al.Regulation role and mechanism of type Ⅳ collagen gene α1 on the proliferation,metastasis and apoptosis of gastric cancer cells[J].Journal of Xinxiang Medical University,2022,39(4):311-317.[doi:10.7683/xxyxyxb.2022.04.003]
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Ⅳ型胶原蛋白基因α1对胃癌细胞增殖、转移及凋亡的调控作用与机制
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
39
期数:
2022年4
页码:
311-317
栏目:
基础研究
出版日期:
2022-04-05

文章信息/Info

Title:
Regulation role and mechanism of type Ⅳ collagen gene α1 on the proliferation,metastasis and apoptosis of gastric cancer cells
作者:
李怡君杨振威陈晓露司望利
(西安市中心医院消化科,陕西 西安 710003)
Author(s):
LI YijunYANG ZhenweiCHEN XiaoluSI Wangli
(Department of Gastroenterology,Xi′an Central Hospital,Xi′an 710003,Shaanxi Province,China)
关键词:
Ⅳ型胶原蛋白基因α1胃癌磷脂酰肌醇3-激酶/蛋白激酶B通路增殖转移
Keywords:
type Ⅳ collagen gene α1gastric cancerphosphoinositide 3-kinase/protein kinase B signalingproliferationmetastasis
分类号:
R735.2
DOI:
10.7683/xxyxyxb.2022.04.003
文献标志码:
A
摘要:
目的 探讨Ⅳ型胶原蛋白基因α1(COL4A1)对胃癌细胞增殖、转移及凋亡的影响以及其潜在的调控机制。方法 采用实时荧光定量聚合酶链式反应(RT-qPCR)法检测胃癌组织和人正常胃黏膜上皮细胞GES-1及胃癌细胞(SGC-7901、MKN-28、BGC-803、MGC-823、MKN-45和AGS)中COL4A1基因的表达。根据6种胃癌细胞中COL4A1基因的表达情况,AGS和SGC-7901细胞被用于后续的细胞功能验证实验。将AGS、SGC-7901细胞接种于6孔板,并于融合度达到90%后,将2种细胞分为对照组、阴性对照(NC)小干扰RNA(siRNA)组、siCOL4A1-1组、siCOL4A1-2组和siCOL4A1-2+740 Y-P组。对照组细胞正常培养,不进行任何处理;NC siRNA组细胞转染NC siRNA,siCOL4A1-1组细胞转染siCOL4A1-1,siCOL4A1-2组细胞转染siCOL4A1-2,siCOL4A1-2+740 Y-P组细胞转染siCOL4A1-2,然后使用50 mg·L-1的740 Y-P处理90 min。采用RT-qPCR检测5组细胞中COL4A1基因的表达情况,根据检测结果选择siCOL4A1-2组细胞进行后续实验。采用细胞计数试剂盒-8检测对照组、NC siRNA组、siCOL4A1-2组和siCOL4A1-2+740 Y-P组AGS和SGC-7901细胞培养1、2、3、4 d时的增殖能力,膜联蛋白-V/异硫氰酸荧光素实验检测4组AGS、SGC-7901细胞凋亡情况,Transwell实验检测4组AGS、SGC-7901细胞迁移和侵袭能力,Western blot检测4组AGS、SGC-7901细胞中磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)通路相关蛋白表达。结果 siCOL4A1-2组AGS细胞的增殖能力在培养3 d和4 d时显著低于对照组和NC siRNA组(P<0.05),SGC-7901细胞的增殖能力在培养1、2、3、4 d时均显著低于对照组和NC siRNA组(P<0.05);siCOL4A1-2+740 Y-P组AGS细胞的增殖能力在培养3 d和4 d时显著高于siCOL4A1-2组(P<0.05),SGC-7901细胞的增殖能力在培养1、2、3、4 d时均显著高于siCOL4A1-2组(P<0.05);其余各组细胞增殖能力比较差异均无统计学意义(P>0.05)。siCOL4A1-2组AGS、SGC-7901细胞的凋亡率均显著高于对照组和NC siRNA组(P<0.05);siCOL4A1-2+740 Y-P组AGS、SGC-7901细胞的凋亡率均显著低于siCOL4A1-2组(P<0.05);其余各组2种细胞凋亡率比较差异均无统计学意义(P>0.05)。siCOL4A1-2组AGS、SGC-7901细胞迁移能力和侵袭能力均显著低于对照组和NC siRNA组(P<0.05);siCOL4A1-2+740 Y-P组AGS、SGC-7901细胞迁移能力和侵袭能力均显著高于siCOL4A1-2组(P<0.05);其余各组2种细胞迁移能力和侵袭能力比较差异无统计学意义(P>0.05)。对照组与NC siRNA组AGS、SGC-7901细胞中p-PI3K/PI3K、p-Akt/Akt值比较差异无统计学意义(P>0.05);siCOL4A1-2组AGS、SGC-7901细胞中p-PI3K/PI3K、p-Akt/Akt值显著低于对照组和NC siRNA组(P<0.01);siCOL4A1-2+740 Y-P组AGS、SGC-7901细胞中p-PI3K/PI3K、p-Akt/Akt值显著高于siCOL4A1-2组(P<0.01)。结论 沉默COL4A1能够通过抑制PI3K/Akt通路的激活抑制胃癌细胞增殖、迁移和侵袭能力,促进细胞凋亡。
Abstract:
Objective To explore the effect of type Ⅳ collagen gene α1 (COL4A1) on the proliferation,metastasis and apoptosis of gastric cancer cells and its potential regulatory mechanism.Methods The expression of COL4A1 gene in gastric cancer tissues,human normal gastric mucosal epithelial cells GES-1 and gastric cancer cells (SGC-7901,MKN-28,BGC-803,mgc-823,MKN-45 and AGS) was detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).According to the expression of COL4A1 gene in six gastric cancer cells,AGS and SGC-7901 cells were used for subsequent cell function verification experiments.AGS and SGC-7901 cells were inoculated into 6-well plates.After the fusion reached 90%,the AGS and SGC-7901 cells were divided into control group,negative control (NC) small interfering RNA (siRNA) group,siCOL4A1-1 group,siCOL4A1-2 group and siCOL4A1-2+740 Y-P group.The cells in the control group were cultured normally without any treatment;the cells in NC siRNA group were transfected with NC siRNA,the cells in siCOL4A1-1 group were transfected with siCOL4A1-1,the cells in siCOL4A1-2 group were transfected with siCOL4A1-2,the cells in siCOL4A1-2+740 Y-P group were transfected with siCOL4A1-2,and then were treated with 740 Y-P (50 mg·L-1) for 90 min.The expression of COL4A1 gene in AGS,SGC-7901 cells in the above five groups was detected by RT-qPCR,and the cells in the siCOL4A1-2 group was selected for follow-up experiments according to the detection results.The proliferation of AGS and SGC-7901 cells in control group,NC siRNA group,siCOL4A1-2 group and siCOL4A1-2+740 Y-P group was detected by cell counting kit-8 at 1,2,3,4 days of culture;the apoptosis of AGS and SGC-7901 cells in the four groups was detected by annexin-V/fluorescein isothiocyanate,the migration and invasion ability of AGS and SGC-7901 cells in the four groups were detected by Transwell experiment;the expression of phosphatidylinositol 3-kinase(PI3K)/protein kinase B(Akt) pathway related protein in AGS and SGC-7901 cells was detected by Western blot.Results The proliferation ability of AGS cells in the siCOL4A1-2 group was significantly lower than that in the control group and the NC siRNA group at 3,4 days of culture (P<0.05);the proliferation ability of SGC-7901 cells in the siCOL4A1-2 group was significantly lower than that in the control group and the NC siRNA group at 1,2,3,4 days of culture (P<0.05).The proliferation ability of AGS cells in the siCOL4A1-2+740 Y-P group was significantly higher than that in the siCOL4A1-2 group at 3,4 days of culture (P<0.05);and the proliferation ability of SGC-7901 cells was significantly higher than that in the siCOL4A1-2 group at 1,2,3 and 4 days of culture (P<0.05).There was no significant difference in the proliferation ability of AGS and SGC-7901 cells among other groups (P>0.05).The apoptosis rates of AGS and SGC-7901 cells in the siCOL4A1-2 group were significantly higher than those in the control group and the NC siRNA group (P<0.05);the apoptosis rates of AGS and SGC-7901 cells in the siCOL4A1-2+740 Y-P group were significantly lower than those in the siCOL4A1-2 group (P<0.05);there was no significant difference in the apoptosis rates of AGS and SGC-7901 cells among other groups (P>0.05).The migration and invasion ability of AGS and SGC-7901 cells in the siCOL4A1-2 group were significantly lower than those in the control group and the NC siRNA group (P<0.05);the migration and invasion ability of AGS and SGC-7901 cells in the siCOL4A1-2+740 Y-P group were significantly higher than those in the siCOL4A1-2 group (P<0.05);there was no significant difference in the migration and invasion ability of AGS and SGC-7901 cells among other groups (P>0.05).There was no significant difference in the values of p-PI3K/PI3K and p-Akt/Akt in AGS and SGC-7901 cells between the control group and the NC siRNA group (P>0.05);the values of p-PI3K/PI3K and p-Akt/Akt in AGS and SGC-7901 cells in siCOL4A1-2 group were significantly lower than those in the control group and the NC siRNA group (P<0.05);the values of p-PI3K/PI3K and p-Akt/Akt in AGS and SGC-7901 cells in siCOL4A1-2+740 Y-P group were significantly higher than those in the siCOL4A1-2 group (P<0.05).Conclusion Silencing COL4A1 can inhibit the proliferation,migration and invasion and promote apoptosis of gastric cancer cells by inhibiting the activation of PI3K/Akt pathway.

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更新日期/Last Update: 2022-04-05