GClnc1通过SIRT3/FOXO3/SOD2信号通路促进胃癌细胞SGC7901的增殖和迁移
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摘要
目的探究lncRNA GClnc1是否通过SIRT3/FOXO3/SOD2调控胃癌细胞增殖和迁移。方法通过转染慢病毒的方法,在人胃癌细胞SGC7901中分别过表达和沉默GClnc1,采用Western blot和qRT-PCR检测SIRT3/FOXO3/SOD2 mRNA和蛋白水平的改变;在过表达GClnc1的同时分别沉默SIRT3/FOXO3/SOD2,采用CCK-8检测细胞增殖能力,划痕实验检测细胞迁移能力。结果过表达GClnc1可以上调SIRT3/FOXO3/SOD2的mRNA和蛋白水平(P<0.05),并增强SGC7901细胞的增殖[(1.00±0.14)vs(1.79±0.21),P<0.05]和迁移能力(10.87±0.76 vs 16.53±2.25,P<0.05),而沉默GClnc1则降低SIRT3/FOXO3/SOD2的mRNA和蛋白水平(P<0.05),并减少细胞增殖(1.00±0.14 vs 0.62±0.13,P<0.05)和迁移能力(10.87±0.76 vs 6.46±0.89,P<0.01)。沉默SIRT3/FOXO3/SOD2可以消除过表达GClnc1对细胞增殖和迁移能力的促进作用。结论 GClnc1通过激活SIRT3/FOXO3/SOD2信号通路促进SGC7901细胞增殖和迁移。
Objective To investigate whether the long noncoding RNA(lncRNA) GClnc1 regulates the proliferation and migration of gastric cancer cells by modulating the expression of SIRT3/FOXO3/SOD2 pathway. Methods We transfected human gastric cancer SGC7901 cells with lentiviral vectors to induce overexpression or knockdown of GClnc1, and examined the changes in the expression of SIRT3/FOXO3/SOD2 at both mRNA and protein levels using qRT-PCR and Western blotting. We also observed the effects of SIRT3/FOXO3/SOD2 silencing on the proliferation and migration of GClnc1-overexpressing SGC7901 cells using CCK8 assay and scratch wound healing assay, respectively. Results Overexpression of GClnc1 significantly up-regulated the mRNA and protein levels of SIRT3/FOXO3/SOD2(P<0.05), and obviously enhanced the proliferation and migration of SGC7901 cells(P<0.05). GClnc1 knockdown significantly lowered the the mRNA and protein levels of SIRT3/FOXO3/SOD2(P<0.05) and inhibited the proliferation and migration of the cells(P<0.05). Silencing SIRT3/FOXO3/SOD2 obviously eliminated the effect of GClnc1 overexpression on the cell proliferation and migration. Conclusion GClnc1 promotes proliferation and migration of SGC7901 cells by activating SIRT3/FOXO3/SOD2 pathway.
Objective To investigate whether the long noncoding RNA(lncRNA) GClnc1 regulates the proliferation and migration of gastric cancer cells by modulating the expression of SIRT3/FOXO3/SOD2 pathway. Methods We transfected human gastric cancer SGC7901 cells with lentiviral vectors to induce overexpression or knockdown of GClnc1, and examined the changes in the expression of SIRT3/FOXO3/SOD2 at both mRNA and protein levels using qRT-PCR and Western blotting. We also observed the effects of SIRT3/FOXO3/SOD2 silencing on the proliferation and migration of GClnc1-overexpressing SGC7901 cells using CCK8 assay and scratch wound healing assay, respectively. Results Overexpression of GClnc1 significantly up-regulated the mRNA and protein levels of SIRT3/FOXO3/SOD2(P<0.05), and obviously enhanced the proliferation and migration of SGC7901 cells(P<0.05). GClnc1 knockdown significantly lowered the the mRNA and protein levels of SIRT3/FOXO3/SOD2(P<0.05) and inhibited the proliferation and migration of the cells(P<0.05). Silencing SIRT3/FOXO3/SOD2 obviously eliminated the effect of GClnc1 overexpression on the cell proliferation and migration. Conclusion GClnc1 promotes proliferation and migration of SGC7901 cells by activating SIRT3/FOXO3/SOD2 pathway.
引文
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[12] COOMANS DE BRACHèNE A, DEMOULIN J B. FOXO transcription factors in cancer development and therapy[J]. Cell Mol Life Sci, 2016, 73(6): 1159-1172. DOI: 10.1007/s00018-015-2112-y.
[13] KIM Y S, GUPTA VALLUR P, PHA TON R, et al. Insights into the dichotomous regulation of SOD2 in cancer[J]. Antioxidants (Basel), 2017, 6(4): E86. DOI: 10.3390/antiox6040086.
[14] LIU Z H, HE Q T, DING X Q, et al. SOD2 is a C-myc target gene that promotes the migration and invasion of tongue squamous cell carcinoma involving cancer stem-like cells[J]. Int J Biochem Cell Biol, 2015, 60: 139-146. DOI: 10.1016/j.biocel.2014.12.022.
[15] HU J, HWANG S S, LIESA M, et al. Antitelomerase therapy provokes ALT and mitochondrial adaptive mechanisms in cancer[J]. Cell, 2012, 148(4): 651-663. DOI: 10.1016/j.cell.2011.12.028.
[16] LIU X H, ZHANG L, WANG P, et al. Sirt3-dependent deacetylation of SOD2 plays a protective role against oxidative stress in oocytes from diabetic mice[J]. Cell Cycle, 2017, 16(13): 1302-1308. DOI: 10.1080/15384101.2017.1320004.
[2] 左婷婷, 郑荣寿, 曾红梅, 等. 中国胃癌流行病学现状[J]. 中国肿瘤临床, 2017, 44(1): 52-58. DOI: 10.3969/j.issn.1000-8179.2017.01.881. ZUO T T, ZHENG R S, ZENG H M, et al. Epidemiology of stomach cancer in China[J]. Chin Clin Oncol, 2017, 44(1): 52-58. DOI: 10.3969/j.issn.1000-8179.2017.01.881.
[3] COHEN J D, LI L, WANG Y X, et al. Detection and localization of surgically resectable cancers with a multi-analyte blood test[J]. Science, 2018, 359(6378): 926-930. DOI: 10.1126/science.aar3247.
[4] SUN T T, HE J, LIANG Q, et al. LncRNA GClnc1 promotes gastric carcinogenesis and may act as a modular scaffold of WDR5 and KAT2A complexes to specify the histone modification pattern[J]. Cancer Discov, 2016, 6(7): 784-801. DOI: 10.1158/2159-8290.CD-15-0921.
[5] KENNY T C, HART P, RAGAZZI M, et al. Selected mitochondrial DNA landscapes activate the SIRT3 axis of the UPRmt to promote metastasis[J]. Oncogene, 2017, 36(31): 4393-4404. DOI: 10.1038/onc.2017.52.
[6] SCHMITT A M, CHANG H Y. Long noncoding RNAs in cancer pathways[J]. Cancer Cell, 2016, 29(4): 452-463. DOI: 10.1016/j.ccell.2016.03.010.
[7] YANG G D, LU X Z, YUAN L J. LncRNA: a link between RNA and cancer[J]. Biochim Biophys Acta, 2014, 1839(11): 1097-1109. DOI: 10.1016/j.bbagrm.2014.08.012.
[8] CHEN Y, FU L L, WEN X, et al. Sirtuin-3 (SIRT3), a therapeutic target with oncogenic and tumor-suppressive function in cancer[J]. Cell Death Dis, 2014, 5: e1047. DOI: 10.1038/cddis.2014.14.
[9] SUNDARESAN N R, SAMANT S A, PILLAI V B, et al. SIRT3 is a stress-responsive deacetylase in cardiomyocytes that protects cells from stress-mediated cell death by deacetylation of Ku70[J]. Mol Cell Biol, 2008, 28(20): 6384-6401. DOI: 10.1128/MCB.00426-08.
[10] ALHAZZAZI T Y, KAMARAJAN P, JOO N, et al. Sirtuin-3 (SIRT3), a novel potential therapeutic target for oral cancer[J]. Cancer, 2011, 117(8): 1670-1678. DOI: 10.1002/cncr.25676.
[11] WEBB A E, BRUNET A. FOXO transcription factors: key regulators of cellular quality control[J]. Trends Biochem Sci, 2014, 39(4): 159-169. DOI: 10.1016/j.tibs.2014.02.003.
[12] COOMANS DE BRACHèNE A, DEMOULIN J B. FOXO transcription factors in cancer development and therapy[J]. Cell Mol Life Sci, 2016, 73(6): 1159-1172. DOI: 10.1007/s00018-015-2112-y.
[13] KIM Y S, GUPTA VALLUR P, PHA TON R, et al. Insights into the dichotomous regulation of SOD2 in cancer[J]. Antioxidants (Basel), 2017, 6(4): E86. DOI: 10.3390/antiox6040086.
[14] LIU Z H, HE Q T, DING X Q, et al. SOD2 is a C-myc target gene that promotes the migration and invasion of tongue squamous cell carcinoma involving cancer stem-like cells[J]. Int J Biochem Cell Biol, 2015, 60: 139-146. DOI: 10.1016/j.biocel.2014.12.022.
[15] HU J, HWANG S S, LIESA M, et al. Antitelomerase therapy provokes ALT and mitochondrial adaptive mechanisms in cancer[J]. Cell, 2012, 148(4): 651-663. DOI: 10.1016/j.cell.2011.12.028.
[16] LIU X H, ZHANG L, WANG P, et al. Sirt3-dependent deacetylation of SOD2 plays a protective role against oxidative stress in oocytes from diabetic mice[J]. Cell Cycle, 2017, 16(13): 1302-1308. DOI: 10.1080/15384101.2017.1320004.
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