小泛素样修饰蛋白1假基因3通过蛋白激酶B信号通路影响非小细胞肺癌细胞系H1299增殖和凋亡

徐亮 杨剑烨 杨飞燕 杨国彪

解剖学报 ›› 2021, Vol. 52 ›› Issue (3) : 410-417.

PDF(9132 KB)
欢迎访问《解剖学报》官方网站!今天是 English
PDF(9132 KB)
解剖学报 ›› 2021, Vol. 52 ›› Issue (3) : 410-417. DOI: 10.16098/j.issn.0529-1356.2021.03.013
细胞和分子生物学

小泛素样修饰蛋白1假基因3通过蛋白激酶B信号通路影响非小细胞肺癌细胞系H1299增殖和凋亡

  • 徐亮 杨剑烨 杨飞燕 杨国彪*
作者信息 +

Effect of small ubiquitin-like modifier 1 pseudogene 3 on the proliferation and apoptosis of non-small cell lung cancer cell line H1299 through protein kinase B signaling pathway

  • XU Liang  YANG Jian-ye  YANG Fei-yan  YANG Guo-biao*
Author information +
文章历史 +

摘要

目的  探讨长链非编码RNA(lncRNA) 小泛素样修饰蛋白1假基因3(SUMO1P3)对非小细胞肺癌细胞增殖和凋亡的影响。   方法  用Real-time PCR方法测定非小细胞肺癌细胞系H1299中SUMO1P3表达变化。在H1299细胞中转染SUMO1P3小干扰RNA(siRNA),Real-time PCR方法测定下调效果, MTT法和5-乙炔基-2’-脱氧尿苷(EdU)法检测细胞增殖, PI单染法测定细胞周期,膜联蛋白 V-FITC(annexin V-FITC)/PI法检测细胞凋亡,TUNEL法检测细胞凋亡,Western blotting方法检测剪切的Caspase-3(c-Caspase-3)、细胞周期蛋白D1(cyclin D1)、P27、磷酸化磷脂酰肌醇-3激酶(p-PI3K)、磷酸化蛋白激酶B(p-Akt)蛋白表达。Akt信号激活剂处理转染SUMO1P3 siRNA后的H1299细胞,同样利用上述方法测定细胞增殖、凋亡和周期变化。样本数均为9。   结果  SUMO1P3在非小细胞肺癌细胞中表达上调。转染SUMO1P3 siRNA后的H1299细胞中SUMO1P3表达水平降低,细胞增殖能力下降,细胞G0/G1期比例升高,细胞凋亡率升高,细胞中c-Caspase-3和P27蛋白水平升高,cyclin D1、p-PI3K和p-Akt蛋白水平下降。Akt信号激活剂可以逆转SUMO1P3 siRNA对H1299细胞增殖抑制、周期阻滞和凋亡促进作用。   结论  下调SUMO1P3抑制非小细胞肺癌H1299细胞增殖并诱导细胞凋亡,作用机制与降低Akt信号通路激活水平有关。

Abstract

Objective  To investigate the effect of long noncoding RNA (lncRNA) small ubiquitin-like modifier 1 pseudogene 3(SUMO1P3) on the proliferation and apoptosis of non-small cell lung cancer cell line 1299.   Methods  Determination of SUMO1P3 expression in non-small cell lung cancer cells by Real-time PCR. SUMO1P3 small interfering RNA(siRNA) was transfected into H1299 cells, the down regulation effect was determined by Real-time PCR.Cell proliferation was measured by MTT, 5-ethynyl-2’-deoxyuridine(EdU) method, the cell cycle was determined by PI single staining, apoptosis was detected by annexin Ⅴ-FITC/PI, detection of apoptosis by TUNEL,Western blotting was used to detect the expression of cleaved Caspase-3(c-Caspase-3), cyclin D1, P27, phosphorylated phospoinositide 3-kinase (p-PI3K)and phosphorylated protein kinase B(p-Akt). Akt signal activator treated H1299 cells transfected with SUMO1P3 siRNA, cell proliferation, apoptosis and cycle change were also measured by the above methods. The number of samples was 9.    Results  SUMO1P3 was up-regulated in non-small cell lung cancer cells. The expression of SUMO1P3 in H1299 cells decreased after transfection with SUMO1P3 siRNA, cell proliferation decreased, the ratio of G0/G1 phase increased, apoptosis rate increased, c-Caspase-3 and P27 protein in the cells increased, the protein levels of cyclin D1, p-PI3K and p-Akt decreased. Akt signal activator could reverse the inhibition of proliferation, cycle arrest and apoptosis of H1299 cells by SUMO1P3 siRNA.    Conclusion  Down-regulation of SUMO1P3 inhibits the proliferation of nonsmall cell lung cancer H1299 cells and induces apoptosis, the mechanism of action is related to the reduction of the activation level of the Akt signaling pathway.

关键词

小泛素样修饰蛋白1假基因3 / 蛋白激酶B / 免疫印迹法

Key words

Small ubiquitin-like modifier 1 pseudogene 3 / Protein kinase B / Western blotting


引用本文

导出引用
徐亮 杨剑烨 杨飞燕 杨国彪. 小泛素样修饰蛋白1假基因3通过蛋白激酶B信号通路影响非小细胞肺癌细胞系H1299增殖和凋亡[J]. 解剖学报. 2021, 52(3): 410-417 https://doi.org/10.16098/j.issn.0529-1356.2021.03.013
XU Liang YANG Jian-ye YANG Fei-yan YANG Guo-biao. Effect of small ubiquitin-like modifier 1 pseudogene 3 on the proliferation and apoptosis of non-small cell lung cancer cell line H1299 through protein kinase B signaling pathway[J]. Acta Anatomica Sinica. 2021, 52(3): 410-417 https://doi.org/10.16098/j.issn.0529-1356.2021.03.013
中图分类号: R734.2   

参考文献

[1] Gandhi L, Rodríguez-Abreu D, Gadgeel S, et al. Pembrolizumab plus chemotherapy in metastatic non-small-cell lung cancer[J]. N Engl J Med, 2018, 378(22): 2078-2092.
[2] Haratani K, Hayashi H, Chiba Y, et al. Association of immune-related adverse events with nivolumab efficacy in non-small-cell lung cancer[J]. JAMA Oncol, 2018, 4(3): 374-378.
[3] Müller Ⅴ, Oliveira-Ferrer L, Steinbach B, et al. Interplay of lncRNA H19/miR-675 and lncRNA NEAT1/miR-204 in breast cancer[J]. Mol Oncol, 2019, 13(5): 1137-1149.
[4] Mansoori Y, Zendehbad Z, Askari A, et al. Breast cancer-linked lncRNA u-eleanor is upregulated in breast of healthy women with lack or short duration of breastfeeding[J]. J Cell Biochem, 2019, 120(6): 9869-9876.
[5] Cao J, Dong R, Jiang L, et al. LncRNA-MM2P identified as a modulator of macrophage M2 polarization [J]. Cancer Immunol Res, 2019, 7(2): 292-305.
[6] Zhou Y, He P, Xie X, et al. Knockdown of SUMO1P3 represses tumor growth and invasion and enhances radiosensitivity in hepatocellular carcinoma[J]. Mol Cell Biochem, 2019, 450(12): 125-134.
[7] Lin H, Guo Q, Lu S, et al. LncRNA SUMO1P3 promotes proliferation and inhibits apoptosis in colorectal cancer by epigenetically silencing CPEB3[J]. Biochem Biophys Res Commun, 2019, 511(2): 239-245.
[8] Tian C, Jin Y, Shi S. Long noncoding RNA SUMO1P3 may promote cell proliferation, migration, and invasion of pancreatic cancer via EMT signaling pathway[J]. Oncol Lett, 2018, 16(5): 6109-6115.
[9] Zhang Y, Li Y, Han L, et al. SUMO1P3 is associated clinical progression and facilitates cell migration and invasion through regulating miR-136 in nonsmall cell lung cancer[J]. Biomed Pharmacother, 2019, 113(5): 108686-108692.
[10] Zhang X, Wu N, Wang J, et al. LncRNA MEG3 inhibits cell proliferation and induces apoptosis in laryngeal cancer via miR-23a/APAF-1 axis[J]. J Cell Mol Med 2019, 23(10): 6708-6719.
[11] Yang Y, Lv X, Fan Q, et al. Analysis of circulating lncRNA expression profiles in patients with diabetes mellitus and diabetic nephropathy: differential expression profile of circulating lncRNA[J]. Clin Nephrol, 2019, 92(1): 25-35.
[12] Zhou J, Zhou Y, Wang C. LncRNA-MIAT regulates fibrosis in hypertrophic cardiomyopathy (HCM) by mediating the expression of miR-29a-3p[J]. J Cell Biochem, 2019, 120(5): 7265-7275.
[13] Yang J, Meng X, Yu Y, et al. LncRNA POU3F3 promotes proliferation and inhibits apoptosis of cancer cells in triple-negative breast cancer by inactivating caspase 9[J]. Biosci Biotechnol Biochem, 2019, 83(6): 1117-1123.
[14] Liu J, Song Z, Feng C, et al. The long non-coding RNA SUMO1P3 facilitates breast cancer progression by negatively regulating miR-320a [J]. Am J Transl Res, 2017,9(12):5594-5602. 
[15] Zhan Y, Liu Y, Wang C, et al. Increased expression of SUMO1P3 predicts poor prognosis and promotes tumor growth and metastasis in bladder cancer[J]. Oncotarget, 2016, 7(13): 16038-16048. 
[16] Wu S, Chen S, Lin N, et al. Long non-coding RNA SUMO1P3 promotes hepatocellular carcinoma progression through activating Wnt/β-catenin signalling pathway by targeting miR-320a[J]. J Cell Mol Med, 2020, 24(5):3108-3116.
[17] Zhou LL, Wang RT, Zhao Y, et al. Effects of picropodophyllin on proliferation and cell cycle of human colorectal cancer HCT-15 cells and its mechanism [J]. Acta Anatomica Sinica, 2020,51 (2): 233-238.(in Chinese)
周丽丽,王瑞婷,赵杨,等. 鬼臼苦素对人结直肠癌HCT-15细胞增殖和周期的影响及其作用机制[J].解剖学报,2020,51(2):233-238.
[18] Wang S, Wang X, Gao Y, et al. RN181 is a tumour suppressor in gastric cancer by regulation of the ERK/MAPK-cyclin D1/CDK4 pathway[J]. J Pathol, 2019, 248(2): 204-216.
[19] Patel P, Tsiperson V, Gottesman SRS, et al. Dual inhibition of CDK4 and CDK2 via targeting p27 tyrosine phosphorylation induces a potent and durable response in breast cancer cells [J]. Mol Cancer Res, 2018, 16(3): 361-377.
[20] Zhou M, Liu X, Li Z, et al. Caspase-3 regulates the migration, invasion and metastasis of colon cancer cells[J]. Int J Cancer, 2018, 143(4): 921-930.
[21] Zhang C, Li C, Wang Y, et al. Chemotherapeutic paclitaxel and cisplatin differentially induce pyroptosis in A549 lung cancer cells via caspase-3/GSDME activation[J]. Apoptosis, 2019, 24 (34): 312-325.
[22] Zhong JL, Zheng L, He H, et al. Advances in research on biological regulation mechanisms related to PI3K/Akt signaling pathway [J]. Genomics and Applied Biology, 2019, 38 (1): 143-147.(in Chinese)
钟佳琳, 郑立, 贺花, 等. PI3K/Akt 信号通路相关的生物学调控机制研究进展[J]. 基因组学与应用生物学, 2019,38 (1): 143-147.
[23] Zarate YA, Boccuto L, Srikanth S, et al. Constitutive activation of the PI3K-Akt pathway and cardiovascular abnormalities in an individual with Kosaki overgrowth syndrome[J]. Am J Med Genet A, 2019, 179(6): 1047-1052.
[24] Tian X, Ji Y, Liang Y, et al. LINC00520 targeting miR-27b-3p regulates OSMR expression level to promote acute kidney injury development through the PI3K/Akt signaling pathway [J]. J Cell Physiol, 2019, 234(8): 14221-14233.
[25] Khan MI, Hamid A, Rath S, et al. Akt inhibition modulates H3K4 demethylase levels in pten-null prostate concer[J]. Mol Cancer Ther, 2019, 18(2): 356-363.
[26] Karlsson E, Veenstra C, G?rsj? J, et al. PTPN2 deficiency along with activation of nuclear Akt predict endocrine resistance in breast cancer[J]. J Cancer Res Clin Oncol, 2019, 145(3): 599-607.

基金

浙江省医药卫生科技计划项目

PDF(9132 KB)

Accesses

Citation

Detail

段落导航
相关文章

/