摘要
目的探索微小RNA-21(miR-21)对胰腺癌MiaPaCa-2细胞侵袭能力的影响和机制。方法选取人类胰腺癌MiaPaCa-2细胞,采用简单随机法分为四组:正常组、miR-21过度表达组、空白对照组和miR-21抑制组。通过慢病毒载体感染MiaPaCa-2细胞,调节miR-21的表达,反转录聚合酶链反应(RT-PCR)验证miR-21的表达;体外侵袭实验(Transwell)检测其侵袭能力;流式细胞蛋白检测和RT-PCR检测上皮-间充质转化(EMT)相关蛋白和基因的表达,包括E-钙黏附蛋白(E-cadherin)、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、N-钙黏附蛋白(N-cadherin)和波形蛋白(Vimentin)。两样本间的比较采用t检验。结果miR-21组中miR-21的表达(1.99±0.17)和MiaPaCa-2细胞侵袭能力(128.00±5.66)高于正常组(0.87±0.12、36.50±0.71),差异有统计学意义(t=9.406、39.315,P<0.01),反之miR-21i组中miR-21的表达(0.50±0.01)和MiaPaCa-2细胞侵袭能力(18.50±3.54)低于对照组(0.99±0.07、36.00±3.54),差异有统计学意义(t=13.021、8.760,P<0.01)。miR-21组中E-cadherin蛋白(42.00±0.82)和基因(0.64±0.04)表达低于正常组蛋白(50.67±1.25)和基因(1.00±0.09),差异有统计学意义(t=0.442、5.653,P<0.05);MMP-2、MMP-9、N-cadherin和Vimentin蛋白(56.00±0.81、63.00±0.82、60.00±1.25、99.33±1.25)和基因(1.24±0.09、1.45±0.14、1.31±0.04、2.42±0.09)高于正常组蛋白(49.67±1.24、51.00±0.81、50.66±1.24、51.00±1.41)和基因(0.99±0.07、0.99±0.05、1.00±0.06、1.01±0.03),差异有统计学意义(t=0.442、14.697、8.018、36.250;t=3.449、4.980、7.414、22.807,P<0.01)。miR-21i组其结果正好相反,E-cadherin蛋白(73.67±1.24)和基因(1.56±0.02)高于对照组蛋白(50.66±0.47)和基因(0.99±0.03),差异有统计学意义(t=24.395、27.253,P<0.05);MMP-2、MMP-9、N-cadherin和Vimentin蛋白(41.00±0.82、44.33±1.25、41.33±1.69、23.67±1.24)和基因(0.67±0.04、0.69±0.03、0.63±0.02、0.34±0.02)低于对照组蛋白(49.67±1.70、52.00±0.82、49.66±0.47、50.00±0.82)和基因(0.85±0.09、0.86±0.07、0.95±0.03、0.97±0.05),差异有统计学意义(t=6.500、7.273、6.682、24.982;t=3.005、3.833、14.774、18.337,P<0.01)。结论miR-21可通过调节EMT相关蛋白的变化,促使胰腺癌细胞的侵袭。
Objective To explore the effects of microRNA-21(miR-21)on the invasive ability of pancreatic cancer MiaPaCa-2 cells and its mechanisms.Methods MiaPaCa-2 cells were randomly divided into four groups by a simple random method:normal group,miR-21 overexpression group,blank control group and miR-21 inhibition group.MiaPaCa-2 cells were infected with lentiviral vectors to increase or decrease the expression of miR-21.The reverse transcription-polymerase chain reaction(RT-PCR)verified the expression of miR-21.In vitro invasion assay(Transwell)was used to detect the invasion ability of MiaPaCa-2 cells.Flow cytometry and RT-PCR were used to detect the expression of epithelial-mesenchymal transformation(EMT)related proteins and genes,including E-cadherin,matrix metalloproteinase-2(MMP-2),MMP-9,N-cadherin and Vimentin.The comparison between the two samples was performed by t test.Results In miR-21 group,the expression of miR-21(1.99±0.17)and the invasion ability of MiaPaCa-2 cells(128.00±5.66)were higher than those in normal group(0.87±0.12,36.50±0.71).The difference was statistically significant(t=9.406,39.315,P<0.01).In miR-21i group,the expression of miR-21(0.50±0.01)and the invasion ability of MiaPaCa-2 cells(18.50±3.54)were lower than those in control group(0.99±0.07,36.00±3.54).The difference was statistically significant(t=13.021,8.760,P<0.01).In miR-21 group,the expression of E-cadherin protein(42.00±0.82)and gene(0.64±0.04)were lower than that in normal group(50.67±1.25,1.00±0.09).The difference was statistically significant(t=0.442,5.653,P<0.05).The expression levels of MMP-2,MMP-9,N-cadherin and Vimentin proteins(56.00±0.81,63.00±0.82,60.00±1.25,99.33±1.25)and genes(1.24±0.09,1.45±0.14,1.31±0.04,2.42±0.09)were higher than those in normal group(49.67±1.24,51.00±0.81,50.66±1.24,51.00±1.41;0.99±0.07,0.99±0.05,1.00±0.06,1.01±0.03).The difference was statistically significant(t=0.442,14.697,8.018,36.250;t=3.449,4.980,7.414,22.807,P<0.01).In miR-21i group,the results were opposite.The expression of E-cadherin protein(73.67±1.24)and gene(1.56±0.02)was higher than that in control group(50.66±0.47,0.99±0.03).The difference was statistically significant(t=24.395,P<0.01;t=27.253,P<0.05).The proteins(41.00±0.82,44.33±1.25,41.33±1.69,23.67±1.24)and genes(0.67±0.04,0.69±0.03,0.63±0.02,0.34±0.02)of MMP-2,MMP-9,N-cadherin and Vimentin were lower than those in control group(49.67±1.70,52.00±0.82,49.66±0.47,50.00±0.82;0.85±0.09,0.86±0.07,0.95±0.03,0.97±0.05).The difference was statistically significant(t=6.500,7.273,6.682,24.982;t=3.005,3.833,14.774,18.337,P<0.01).Conclusion MiR-21 can promote the invasion of pancreatic cancer cells by regulating the changes of EMT-related proteins.
作者
王闯
周晓华
姚金科
谭硕果
梁永艺
Wang Chuang;Zhou Xiaohua;Yao Jinke;Tan Shuoguo;Liang Yongyi(Department of Hepatobiliary Surgery,the Fourth Affiliated Hospital of Guangzhou Medical University,Guangzhou 511300,China;Department of Ultrasound,the Second Affiliated Hospital of Guangzhou Medical University,Guangzhou 510260,China)
出处
《中华实验外科杂志》
CAS
北大核心
2023年第9期1773-1776,共4页
Chinese Journal of Experimental Surgery
基金
广州市增城区科技创新资金计划项目(增科工商信合同(2021)056)。
关键词
胰腺癌
微小RNA
侵袭
上皮-间充质转化
Pancreatic cancer
MicroRNA
Invasion
Epithelial-mesenchymal transformation
