摘要
目的:探讨长链非编码RNA(long chain non-coding RNA,lnc RNA)肺腺癌转移相关转录因子1(metastasis-associated lung adenocarcinoma transcript 1,MALAT1)通过调控miR-204的表达影响胰腺癌细胞恶性生物学行为的作用及其可能机制。方法:收集2016年10月至2016年12月在河南省中医院6例胰腺癌手术切除标本及其对应的癌旁组织标本。用实时荧光定量PCR检测胰腺癌组织和癌旁组织、5种胰腺癌细胞(Bx PC-3、HS-7667、PANC-1、As PC-1和SW-1990)中lnc RNA MALAT1的表达,双荧光素酶报告基因检测MALAT1与miR-204的相互作用,流式细胞术分析MALAT-1对胰腺癌细胞的细胞周期及细胞凋亡的影响;划痕愈合实验和Transwell侵袭实验检测MALAT1和miR-204对胰腺癌细胞迁移和侵袭能力的影响,Western blotting检测MALAT1对上皮间质转化相关蛋白的影响。结果:lnc RNA MALAT1在胰腺癌组织中表达水平明显高于癌旁组织(1.85±0.52vs 0.34±0.12,P<0.05),MALAT1在SW-1990细胞中的表达水平最高(P<0.05)。lnc RNA MALAT1能与miR-204的3’UTR特异性结合,调控miR-204的表达活性。抑制MALAT1表达后,可诱导SW-1990细胞G2/M细胞周期停滞、促进细胞凋亡,SW-1990细胞的迁移和侵袭能力减弱(均P<0.05),下调SW-1990细胞N-cadherin、E-cadherin和vimentin的表达。过表达miR-204可促进SW-1990细胞迁移和侵袭。结论:lnc RNA MALAT1通过靶向调控miR-204表达影响胰腺癌细胞的恶性生物学行为,在胰腺癌的发生发展过程中起重要作用。
Objective： To investigate the effect of long-chain non-coding RNA metastasis-associated lung adenocarcinoma transcript 1 （lncRNA MALAT1 ） on the malignant biological behavior of pancreatic cancer cells by regulating miR-204 expression and its possible mechanisms. Methods： Six pairs of pancreatic cancer tissues and corresponding para-cancerous tissues were sampled during surgery from October 2016 to December 2016 in Henan Provincial Hospital of Traditional Chinese Medicine. qPCR was used to detect the ex- pression of lncRNA MALAT1 in pancreatic cancer tissues, adjacent normal tissues and different pancreatic cancer cells （BxPC-3, HS- 7667, PANC-1, AsPC-1 and SW-1990）. Double luciferase reporter gene assay was used to detect the interaction between MALAT 1 and miR-204. Flow cytometry was used to determine the effect of MALAT1 on cell cycle and apoptosis of pancreatic cancer cells. The ef- fects of MALAT1 and miR-204 on the migration and invasion of pancreatic cancer cells were examined by Wound-healing assay and Transwell invasion assay, respectively. The effect of MALAT1 on EMT-related proteins was detected by Western blotting. Results： Compared with para-cancer tissue, the expression of lncRNA MALAT1 in pancreatic cancer tissues was significantly increased （1.85±0.52 vs 0.34±0.12, P〈0.05）. The expression level of lncRNA MALAT1 in pancreatic cancer SW-1990 cell line was the highest （P〈 0.05）. lncRNA MALAT1 could bind specifically to the 3 ＇UTR of miR-204 to modulate the expression of miR-204. Inhibition of MALAT1 expression could induce G2/M cell cycle arrest, and thus promote SW-1990 cell apoptosis; moreover, the ability of migration and invasion of SW-1990 cells was weakened and EMT-related proteins （N-cadherin, E-cadherin and vimentin） were down-regulated af- ter inhibiting the expression of MALAT1. However, over-expression of miR-204 could promote the migration and invasion of SW-1990 Cells. Conclusion： lncRNA MALAT1 plays an important role in the development and progression of pancreatic cancer. It can regulate the malignant biological behavior of pancreatic cancer cells by targeting miR-204.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2018年第1期79-84,共6页
Chinese Journal of Cancer Biotherapy
基金
河南省科技攻关项目资助(No.142102310040)~~
