山东医药
山東醫藥
산동의약
SHANDONG MEDICAL JOURNAL
2015年
22期
12-15
,共4页
孙硕文%朱之燕%杜玮%王豪%刘喆
孫碩文%硃之燕%杜瑋%王豪%劉喆
손석문%주지연%두위%왕호%류철
非小细胞肺癌%SIRT1-shRNA慢病毒表达载体%SIRT1基因%E-cadherin蛋白%β-catenin蛋白%细胞黏附%细胞迁移
非小細胞肺癌%SIRT1-shRNA慢病毒錶達載體%SIRT1基因%E-cadherin蛋白%β-catenin蛋白%細胞黏附%細胞遷移
비소세포폐암%SIRT1-shRNA만병독표체재체%SIRT1기인%E-cadherin단백%β-catenin단백%세포점부%세포천이
non-small-cell lung cancer ( NSCLC)%SIRT1-shRNA lentiviral expression vector%SIRT1 gene%E-cad-herin protein%β-catenin protein%cell adhesion%cell migration
目的：探讨SIRT1-shRNA慢病毒表达载体转染对人非小细胞肺癌A549细胞SIRT1基因表达、黏附能力、迁移能力及E-cadherin、β-catenin mRNA和蛋白表达量的影响。方法构建SIRT1-shRNA慢病毒表达载体，并用其转染A549细胞。对照组转染277空载体，观察组转染277-iSIRT1。用RT-PCR法检测两组细胞SIRT1 mRNA表达，用细胞黏附实验检测两组细胞黏附能力，划痕实验检测细胞迁移能力，RT-PCR法检测细胞内E-cadherin、β-catenin mRNA表达，Western blot法检测细胞内E-cadherin、β-catenin蛋白表达。结果观察组、对照组A549细胞SIRT1 mRNA分别为0．48±0．26、1．00±0．00（P＜0．05），黏附细胞数分别为（196．6±9．8）、（125．6±9．8）个（P＜0．05），细胞相对迁移率分别为65％±2％、100％±0（P＜0．05），E-cadherin蛋白表达量分别为1．27±0．16、1．00±0．00（P＜0．05），β-catenin蛋白表达量分别为1．24±0．13、1．00±0．00（P＜0．05）。结论 SIRT1-shRNA慢病毒表达载体可明显下调A549细胞SIRT1表达，提高其下游基因E-cadherin、β-catenin mRNA和蛋白表达量，增强细胞黏附能力，降低细胞迁移能力。
目的：探討SIRT1-shRNA慢病毒錶達載體轉染對人非小細胞肺癌A549細胞SIRT1基因錶達、黏附能力、遷移能力及E-cadherin、β-catenin mRNA和蛋白錶達量的影響。方法構建SIRT1-shRNA慢病毒錶達載體，併用其轉染A549細胞。對照組轉染277空載體，觀察組轉染277-iSIRT1。用RT-PCR法檢測兩組細胞SIRT1 mRNA錶達，用細胞黏附實驗檢測兩組細胞黏附能力，劃痕實驗檢測細胞遷移能力，RT-PCR法檢測細胞內E-cadherin、β-catenin mRNA錶達，Western blot法檢測細胞內E-cadherin、β-catenin蛋白錶達。結果觀察組、對照組A549細胞SIRT1 mRNA分彆為0．48±0．26、1．00±0．00（P＜0．05），黏附細胞數分彆為（196．6±9．8）、（125．6±9．8）箇（P＜0．05），細胞相對遷移率分彆為65％±2％、100％±0（P＜0．05），E-cadherin蛋白錶達量分彆為1．27±0．16、1．00±0．00（P＜0．05），β-catenin蛋白錶達量分彆為1．24±0．13、1．00±0．00（P＜0．05）。結論 SIRT1-shRNA慢病毒錶達載體可明顯下調A549細胞SIRT1錶達，提高其下遊基因E-cadherin、β-catenin mRNA和蛋白錶達量，增彊細胞黏附能力，降低細胞遷移能力。
목적：탐토SIRT1-shRNA만병독표체재체전염대인비소세포폐암A549세포SIRT1기인표체、점부능력、천이능력급E-cadherin、β-catenin mRNA화단백표체량적영향。방법구건SIRT1-shRNA만병독표체재체，병용기전염A549세포。대조조전염277공재체，관찰조전염277-iSIRT1。용RT-PCR법검측량조세포SIRT1 mRNA표체，용세포점부실험검측량조세포점부능력，화흔실험검측세포천이능력，RT-PCR법검측세포내E-cadherin、β-catenin mRNA표체，Western blot법검측세포내E-cadherin、β-catenin단백표체。결과관찰조、대조조A549세포SIRT1 mRNA분별위0．48±0．26、1．00±0．00（P＜0．05），점부세포수분별위（196．6±9．8）、（125．6±9．8）개（P＜0．05），세포상대천이솔분별위65％±2％、100％±0（P＜0．05），E-cadherin단백표체량분별위1．27±0．16、1．00±0．00（P＜0．05），β-catenin단백표체량분별위1．24±0．13、1．00±0．00（P＜0．05）。결론 SIRT1-shRNA만병독표체재체가명현하조A549세포SIRT1표체，제고기하유기인E-cadherin、β-catenin mRNA화단백표체량，증강세포점부능력，강저세포천이능력。
Objective To investigate effect of SIRT1-shRNA lentiviral expression vector transfection on the cell adhe-sion, migration and the expression of E-cadherin and β-catenin in non-small-cell lung cancer ( NSCLC) A549 cell line. Methods The SIRT1-shRNA lentiviral expression vector was constructed and we used it to transfect the A549 cells.The cells infected with 277 empty vector were used as the control group and the cells infected with 277-iSIRT1 were used as the observation group.The SIRT1 mRNA expression in the two groups was detected by RT-PCR.Cell adhesion status of A549 cells in the two groups was detected by cell adhesion assay, cell migration was detected by wound healing assay, and the protein and mRNA expression levels of E-cadherin andβ-catenin were detected by RT-PCR and Western blotting, respec-tively.Results the SIRT1 mRNA of A549 cells in the observation group and control group were respectively 0.48 ±0.26 and 1.00 ±0.00 (P<0.05), the adhesive cells of the observation group and control group were 196.6 ±9.8 and 125.6 ±9.8, respectively (P<0.05), the relative migration rates of the observation group and control group were 0.65 ±0.02 and 1.00 ±0.00, respectively (P<0.05).and the E-cadherin expression levels of the observation group and control group were 1.27 ±0.16 and 1.00 ±0.00 (P<0.05), andβ-catenin expression levels were 1.24 ±0.13 and 1.00 ±0.00 (P<0.05).Conclusion SIRT1-shRNA lentiviral expression vector may significantly down-regulate the expression of SIRT1 in A549 cells, increase the expression levels of E-cadherin andβ-catenin mRNA and protein, enhance the capacity of cell ad-hesion and reduce the cell migration ability.
