姜黄素调节miR-133a表达对肝癌细胞迁移和侵袭的影响
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摘要
背景姜黄素对包括肝癌在内的多种肿瘤的发生发展表现出较好的抑制作用,但其抗肝癌的作用机制尚不完全清晰.有研究发现,姜黄素可通过调控miR-133a表达抑制胃癌细胞增殖和转移; miR-133a在肝癌中低表达的结果已有数据证实,但姜黄素是否介导miR-133a表达发挥抗肝癌的作用并不清楚.目的探讨姜黄素调节miR-133a表达对肝癌细胞迁移和侵袭的影响.方法以姜黄素(0、10、20μmol/L)处理肝癌SMMC-7721细胞48 h后, MTT法检测细胞活力, Transwell小室检测细胞的迁移和侵袭; RT-PCR检测细胞中miR-133a表达.采用RT-PCR检测正常肝LO2细胞和肝癌SMMC-7721细胞中miR-133a表达;将miR-133a模拟物转染至SMMC-7721细胞后, Transwell小室检测miR-133a对姜黄素作用前后细胞迁移和侵袭的影响.结果姜黄素能够有效抑制SMMC-7721细胞活力(10μmol/L姜黄素存活率:0.71±0.07 vs 1.02±0.09; 20μmol/L姜黄素存活率:0.45±0.05 vs 1.02±0.09)、迁移(52.32±5.48 vs121.43±12.35)和侵袭(46.33±5.38 vs109.25±10.75),上调miR-133a表达(10μmol/L姜黄素:1.62±0.11 vs 1.00±0.09; 20μmol/L姜黄素:2.96±0.25vs1.00±0.09);与LO2细胞相比,SMCC-7721细胞中miR-133a的表达水平明显降低(0.32±0.03 vs1.03±0.08);上调miR-133a表达后,SMCC-7721细胞的迁移(32.84±3.95 vs 96.35±9.08)和侵袭(42.75±5.06vs119.32±11.71)能力均明显减弱,同时姜黄素对SMCC-7721细胞迁移(29.6±3.32 vs134.62±13.41)和侵袭(31.86±4.05 vs 129.73±12.74)的抑制作用增强.结论姜黄素可通过上调miR-133a表达抑制肝癌细胞的迁移和侵袭.
BACKGROUND Curcumin has a good inhibitory effect on the occurrence and development of many kinds of tumors,including hepatocellular carcinoma, but its antihepatocellular carcinoma mechanism is not completely clear.Some studies have found that curcumin can inhibit the proliferation and metastasis of gastric cancer cells by regulating the expression of miR-133a.The low expression of miR-133a in hepatocellular carcinoma has been confirmed by data, but whether curcumin regulated expression of miR-133a plays an anti-hepatocellular carcinoma role is not clear.AIM To investigate the effect of curcumin on the migration and invasion of hepatocellular carcinoma cells and explore the underlying mechanism by detecting the expression of miR-133a.METHODS After treatment of liver cancer SMMC-7721 cells with curcumin(0, 10, and 20 μmol/L) for 48 h, cell viability was detected by MTT assay, cell migration and invasion were measured by transwell assay, and the expression of miR-133a in the cells was detected by RT-PCR.The expression of miR-133a in normal liver LO2 cells and hepatocellular carcinoma SMMC-7721 cells was detected by RT-PCR.After transfection with miR-133a analogue to SMMC-7721 cells, the effect of miR-133a on migration and invasion of cells before and after curcumin treatment was detected by transwell assay.RESULTS Curcumin effectively inhibited SMMC-7721 cell viability(10 μmol/L curcumin: 0.71 + 0.07 vs 1.02 + 0.09; 20 μmol/L: 0.45 + 0.05 vs 1.02 + 0.09), migration(52.32 ± 5.48 vs 121.43 ± 12.35), and invasion(46.33 ± 5.38 vs 109.25 ± 10.75) and increased miR-133a expression(10 μmol/L curcumin: 1.62 ± 0.11 vs 1.00 ± 0.09; 20 μmol/L: 2.96 ± 0.25 vs 1.00 ± 0.09).Compared with LO2 cells, the expression level of miR-133a(0.32 ± 0.03 vs 1.03 ± 0.08) in SMCC-7721 cells was decreased obviously.After increasing the expression of miR-133a, the migration(32.84 ± 3.95 vs 96.35 ± 9.08) and invasion(42.75 ± 5.06 vs 119.32 ± 11.71) of SMCC-7721 cells were significantly decreased, and the inhibitory effect of curcumin on migration(29.6 ± 3.32 vs 134.62 ± 13.41) and invasion(31.86 ± 4.05 vs 129.73 ± 12.74) of SMCC-7721 cells was enhanced.CONCLUSION Curcumin can inhibit the migration and invasion of hepatoma cells by up-regulating the expression of miR-133a.
BACKGROUND Curcumin has a good inhibitory effect on the occurrence and development of many kinds of tumors,including hepatocellular carcinoma, but its antihepatocellular carcinoma mechanism is not completely clear.Some studies have found that curcumin can inhibit the proliferation and metastasis of gastric cancer cells by regulating the expression of miR-133a.The low expression of miR-133a in hepatocellular carcinoma has been confirmed by data, but whether curcumin regulated expression of miR-133a plays an anti-hepatocellular carcinoma role is not clear.AIM To investigate the effect of curcumin on the migration and invasion of hepatocellular carcinoma cells and explore the underlying mechanism by detecting the expression of miR-133a.METHODS After treatment of liver cancer SMMC-7721 cells with curcumin(0, 10, and 20 μmol/L) for 48 h, cell viability was detected by MTT assay, cell migration and invasion were measured by transwell assay, and the expression of miR-133a in the cells was detected by RT-PCR.The expression of miR-133a in normal liver LO2 cells and hepatocellular carcinoma SMMC-7721 cells was detected by RT-PCR.After transfection with miR-133a analogue to SMMC-7721 cells, the effect of miR-133a on migration and invasion of cells before and after curcumin treatment was detected by transwell assay.RESULTS Curcumin effectively inhibited SMMC-7721 cell viability(10 μmol/L curcumin: 0.71 + 0.07 vs 1.02 + 0.09; 20 μmol/L: 0.45 + 0.05 vs 1.02 + 0.09), migration(52.32 ± 5.48 vs 121.43 ± 12.35), and invasion(46.33 ± 5.38 vs 109.25 ± 10.75) and increased miR-133a expression(10 μmol/L curcumin: 1.62 ± 0.11 vs 1.00 ± 0.09; 20 μmol/L: 2.96 ± 0.25 vs 1.00 ± 0.09).Compared with LO2 cells, the expression level of miR-133a(0.32 ± 0.03 vs 1.03 ± 0.08) in SMCC-7721 cells was decreased obviously.After increasing the expression of miR-133a, the migration(32.84 ± 3.95 vs 96.35 ± 9.08) and invasion(42.75 ± 5.06 vs 119.32 ± 11.71) of SMCC-7721 cells were significantly decreased, and the inhibitory effect of curcumin on migration(29.6 ± 3.32 vs 134.62 ± 13.41) and invasion(31.86 ± 4.05 vs 129.73 ± 12.74) of SMCC-7721 cells was enhanced.CONCLUSION Curcumin can inhibit the migration and invasion of hepatoma cells by up-regulating the expression of miR-133a.
引文
1 Fu H,Wang C,Yang D,Wei Z,Xu J,Hu Z,Zhang Y,Wang W,Yan R,Cai Q.Curcumin regulates proliferation,autophagy,and apoptosis in gastric cancer cells by affecting PI3K and P53signaling.J Cell Physiol 2018;233:4634-4642[PMID:28926094DOI:10.1002/jcp.26190]
2 Zheng BZ,Liu TD,Chen G,Zhang JX,Kang X.The effect of curcumin on cell adhesion of human esophageal cancer cell.Eur Rev Med Pharmacol Sci 2018;22:551-560[PMID:29424917DOI:10.26355/eurrev_201801_14209]
3 Wang R,Li J,Zhao Y,Li Y,Yin L.Investigating the therapeutic potential and mechanism of curcumin in breast cancer based on RNA sequencing and bioinformatics analysis.Breast Cancer 2018;25:206-212[PMID:29139094 DOI:10.1007/s12282-017-0816-6]
4邵娟.姜黄素调节miR-133a-3p靶向MMP15干预胃癌增殖、转移的作用及机制研究.扬州大学,2017
5 Liang HW,Yang X,Wen DY,Gao L,Zhang XY,Ye ZH,Luo J,Li ZY,He Y,Pang YY,Chen G.Utility of miR-133a-3p as a diagnostic indicator for hepatocellular carcinoma:An investigation combined with GEO,TCGA,meta-analysis and bioinformatics.Mol Med Rep 2018;17:1469-1484[PMID:29138825 DOI:10.3892/mmr.2017.8040]
6 Kim HJ,Park SY,Park OJ,Kim YM.Curcumin suppresses migration and proliferation of Hep3B hepatocarcinoma cells through inhibition of the Wnt signaling pathway.Mol Med Rep 2013;8:282-286[PMID:23723038 DOI:10.3892/mmr.2013.1497]
7 Warzyszyńska K,Jonas M,Wasiak D,Kosieradzki M,Ma?kowski P.Accelerated hepatocellular carcinoma recurrence rate after postoperative direct-acting antivirals treatmentpreliminary report.Clin Exp Hepatol 2017;3:194-197[PMID:29255807 DOI:10.5114/ceh.2017.71483]
8 Zhou YM,Zhang XF,Li B,Sui CJ,Yang JM.Postoperative complications affect early recurrence of hepatocellular carcinoma after curative resection.BMC Cancer 2015;15:689[PMID:26466573 DOI:10.1186/s12885-015-1720-0]
9 Yang J,Wang C,Zhang Z,Chen X,Jia Y,Wang B,Kong T.Curcumin inhibits the survival and metastasis of prostate cancer cells via the Notch-1 signaling pathway.APMIS 2017;125:134-140[PMID:28120490 DOI:10.1111/apm.12650]
10 Cao L,Xiao X,Lei J,Duan W,Ma Q,Li W.Curcumin inhibits hypoxia-induced epithelial-mesenchymal transition in pancreatic cancer cells via suppression of the hedgehog signaling pathway.Oncol Rep 2016;35:3728-3734[PMID:27035865 DOI:10.3892/or.2016.4709]
11 Tong W,Wang Q,Sun D,Suo J.Curcumin suppresses colon cancer cell invasion via AMPK-induced inhibition of NF-κB,u PA activator and MMP9.Oncol Lett 2016;12:4139-4146[PMID:27895783 DOI:10.3892/ol.2016.5148]
12 Zhang HH,Zhang Y,Cheng YN,Gong FL,Cao ZQ,Yu LG,Guo XL.Metformin incombination with curcumin inhibits the growth,metastasis,and angiogenesis of hepatocellular carcinoma in vitro and in vivo.Mol Carcinog 2018;57:44-56[PMID:28833603 DOI:10.1002/mc.22718]
13邱伟,刘阳,曹卫,徐军.姜黄素抑制肝癌细胞系HepG2的增殖、侵袭及其机制.现代肿瘤医学2015;23:3221-3225[DOI:10.3969/j.issn.1672-4992.2015.22.004]
14 Wu HJ,Zhuo Y,Zhou YC,Wang XW,Wang YP,Si CY,Wang XH.miR-29a promotes hepatitis B virus replication and expression by targeting SMARCE1 in hepatoma carcinoma.World J Gastroenterol 2017;23:4569-4578[PMID:28740345 DOI:10.3748/wjg.v23.i25.4569]
15 Wang X,Liao Z,Bai Z,He Y,Duan J,Wei L.MiR-93-5p Promotes Cell Proliferation through Down-Regulating PPARGC1A in Hepatocellular Carcinoma Cells by Bioinformatics Analysis and Experimental Verification.Genes(Basel)2018;9:[PMID:29361788 DOI:10.3390/genes9010051]
16周艳华,茅慧,张鹏飞,戴广海.miR-451对肝癌细胞增殖、糖酵解及相关基因表达的影响.现代中西医结合杂志2018;27:934-937[DOI:10.3969/j.issn.1008-8849.2018.09.006]
17 He RQ,Wu PR,Xiang XL,Yang X,Liang HW,Qiu XH,Yang LH,Peng ZG,Chen G.Downregulated miR-23b-3p expression acts as a predictor of hepatocellular carcinoma progression:A study based on public data and RT-qPCR verification.Int JMol Med 2018;41:2813-2831[PMID:29484429 DOI:10.3892/ijmm.2018.3513]
18 Li X,Song H,Liu Z,Bi Y.miR-1260b promotes cell migration and invasion of hepatocellular carcinoma by targeting the regulator of G-protein signaling 22.Biotechnol Lett 2018;40:57-62[PMID:29038925 DOI:10.1007/s10529-017-2455-6]
19 Sui Y,Zhang X,Yang H,Wei W,Wang M.MicroRNA-133a acts as a tumour suppressor in breast cancer through targeting LASP1.Oncol Rep 2018;39:473-482[PMID:29207145 DOI:10.3892/or.2017.6114]
20 Qiu T,Zhou X,Wang J,Du Y,Xu J,Huang Z,Zhu W,Shu Y,Liu P.MiR-145,miR-133a and miR-133b inhibit proliferation,migration,invasion and cell cycle progression via targeting transcription factor Sp1 in gastric cancer.FEBS Lett 2014;588:1168-1177[PMID:24613927 DOI:10.1016/j.febslet.2014.02.054]
21 Wang Y,Li J,Chen H,Mo Y,Ye H,Luo Y,Guo K,Mai Z,Zhang Y,Chen B,Zhou Y,Yang Z.Down-regulation of miR-133a as a poor prognosticator in non-small cell lung cancer.Gene 2016;591:333-337[PMID:27282282 DOI:10.1016/j.gene.2016.06.001]
22 Zhang W,Liu K,Liu S,Ji B,Wang Y,Liu Y.MicroRNA-133a functions as a tumor suppressor by targeting IGF-1Rin hepatocellular carcinoma.Tumour Biol 2015;36:9779-9788[PMID:26156803 DOI:10.1007/s13277-015-3749-8]
23 Cao MT,Liu HF,Liu ZG,Xiao P,Chen JJ,Tan Y,Jiang XX,Jiang ZC,Qiu Y,Huang HJ,Zhang QG,Jiang GM.Curcumin downregulates the expression of Snail via suppressing Smad2pathway to inhibit TGF-β1-induced epithelial-mesenchymal transitions in hepatoma cells.Oncotarget 2017;8:108498-108508[PMID:29312546 DOI:10.18632/oncotarget.22590]
2 Zheng BZ,Liu TD,Chen G,Zhang JX,Kang X.The effect of curcumin on cell adhesion of human esophageal cancer cell.Eur Rev Med Pharmacol Sci 2018;22:551-560[PMID:29424917DOI:10.26355/eurrev_201801_14209]
3 Wang R,Li J,Zhao Y,Li Y,Yin L.Investigating the therapeutic potential and mechanism of curcumin in breast cancer based on RNA sequencing and bioinformatics analysis.Breast Cancer 2018;25:206-212[PMID:29139094 DOI:10.1007/s12282-017-0816-6]
4邵娟.姜黄素调节miR-133a-3p靶向MMP15干预胃癌增殖、转移的作用及机制研究.扬州大学,2017
5 Liang HW,Yang X,Wen DY,Gao L,Zhang XY,Ye ZH,Luo J,Li ZY,He Y,Pang YY,Chen G.Utility of miR-133a-3p as a diagnostic indicator for hepatocellular carcinoma:An investigation combined with GEO,TCGA,meta-analysis and bioinformatics.Mol Med Rep 2018;17:1469-1484[PMID:29138825 DOI:10.3892/mmr.2017.8040]
6 Kim HJ,Park SY,Park OJ,Kim YM.Curcumin suppresses migration and proliferation of Hep3B hepatocarcinoma cells through inhibition of the Wnt signaling pathway.Mol Med Rep 2013;8:282-286[PMID:23723038 DOI:10.3892/mmr.2013.1497]
7 Warzyszyńska K,Jonas M,Wasiak D,Kosieradzki M,Ma?kowski P.Accelerated hepatocellular carcinoma recurrence rate after postoperative direct-acting antivirals treatmentpreliminary report.Clin Exp Hepatol 2017;3:194-197[PMID:29255807 DOI:10.5114/ceh.2017.71483]
8 Zhou YM,Zhang XF,Li B,Sui CJ,Yang JM.Postoperative complications affect early recurrence of hepatocellular carcinoma after curative resection.BMC Cancer 2015;15:689[PMID:26466573 DOI:10.1186/s12885-015-1720-0]
9 Yang J,Wang C,Zhang Z,Chen X,Jia Y,Wang B,Kong T.Curcumin inhibits the survival and metastasis of prostate cancer cells via the Notch-1 signaling pathway.APMIS 2017;125:134-140[PMID:28120490 DOI:10.1111/apm.12650]
10 Cao L,Xiao X,Lei J,Duan W,Ma Q,Li W.Curcumin inhibits hypoxia-induced epithelial-mesenchymal transition in pancreatic cancer cells via suppression of the hedgehog signaling pathway.Oncol Rep 2016;35:3728-3734[PMID:27035865 DOI:10.3892/or.2016.4709]
11 Tong W,Wang Q,Sun D,Suo J.Curcumin suppresses colon cancer cell invasion via AMPK-induced inhibition of NF-κB,u PA activator and MMP9.Oncol Lett 2016;12:4139-4146[PMID:27895783 DOI:10.3892/ol.2016.5148]
12 Zhang HH,Zhang Y,Cheng YN,Gong FL,Cao ZQ,Yu LG,Guo XL.Metformin incombination with curcumin inhibits the growth,metastasis,and angiogenesis of hepatocellular carcinoma in vitro and in vivo.Mol Carcinog 2018;57:44-56[PMID:28833603 DOI:10.1002/mc.22718]
13邱伟,刘阳,曹卫,徐军.姜黄素抑制肝癌细胞系HepG2的增殖、侵袭及其机制.现代肿瘤医学2015;23:3221-3225[DOI:10.3969/j.issn.1672-4992.2015.22.004]
14 Wu HJ,Zhuo Y,Zhou YC,Wang XW,Wang YP,Si CY,Wang XH.miR-29a promotes hepatitis B virus replication and expression by targeting SMARCE1 in hepatoma carcinoma.World J Gastroenterol 2017;23:4569-4578[PMID:28740345 DOI:10.3748/wjg.v23.i25.4569]
15 Wang X,Liao Z,Bai Z,He Y,Duan J,Wei L.MiR-93-5p Promotes Cell Proliferation through Down-Regulating PPARGC1A in Hepatocellular Carcinoma Cells by Bioinformatics Analysis and Experimental Verification.Genes(Basel)2018;9:[PMID:29361788 DOI:10.3390/genes9010051]
16周艳华,茅慧,张鹏飞,戴广海.miR-451对肝癌细胞增殖、糖酵解及相关基因表达的影响.现代中西医结合杂志2018;27:934-937[DOI:10.3969/j.issn.1008-8849.2018.09.006]
17 He RQ,Wu PR,Xiang XL,Yang X,Liang HW,Qiu XH,Yang LH,Peng ZG,Chen G.Downregulated miR-23b-3p expression acts as a predictor of hepatocellular carcinoma progression:A study based on public data and RT-qPCR verification.Int JMol Med 2018;41:2813-2831[PMID:29484429 DOI:10.3892/ijmm.2018.3513]
18 Li X,Song H,Liu Z,Bi Y.miR-1260b promotes cell migration and invasion of hepatocellular carcinoma by targeting the regulator of G-protein signaling 22.Biotechnol Lett 2018;40:57-62[PMID:29038925 DOI:10.1007/s10529-017-2455-6]
19 Sui Y,Zhang X,Yang H,Wei W,Wang M.MicroRNA-133a acts as a tumour suppressor in breast cancer through targeting LASP1.Oncol Rep 2018;39:473-482[PMID:29207145 DOI:10.3892/or.2017.6114]
20 Qiu T,Zhou X,Wang J,Du Y,Xu J,Huang Z,Zhu W,Shu Y,Liu P.MiR-145,miR-133a and miR-133b inhibit proliferation,migration,invasion and cell cycle progression via targeting transcription factor Sp1 in gastric cancer.FEBS Lett 2014;588:1168-1177[PMID:24613927 DOI:10.1016/j.febslet.2014.02.054]
21 Wang Y,Li J,Chen H,Mo Y,Ye H,Luo Y,Guo K,Mai Z,Zhang Y,Chen B,Zhou Y,Yang Z.Down-regulation of miR-133a as a poor prognosticator in non-small cell lung cancer.Gene 2016;591:333-337[PMID:27282282 DOI:10.1016/j.gene.2016.06.001]
22 Zhang W,Liu K,Liu S,Ji B,Wang Y,Liu Y.MicroRNA-133a functions as a tumor suppressor by targeting IGF-1Rin hepatocellular carcinoma.Tumour Biol 2015;36:9779-9788[PMID:26156803 DOI:10.1007/s13277-015-3749-8]
23 Cao MT,Liu HF,Liu ZG,Xiao P,Chen JJ,Tan Y,Jiang XX,Jiang ZC,Qiu Y,Huang HJ,Zhang QG,Jiang GM.Curcumin downregulates the expression of Snail via suppressing Smad2pathway to inhibit TGF-β1-induced epithelial-mesenchymal transitions in hepatoma cells.Oncotarget 2017;8:108498-108508[PMID:29312546 DOI:10.18632/oncotarget.22590]