Study on the mechanism of miR-515-5p enhanced the cisplatin sensitivity of NSCLC A549/DDP cells via targeting RING1

doi:10.3877/cma.j.issn.1674-6902.2021.05.003

Abstract

Abstract:

Objective

To investigate the effect and mechanism of microRNA-515-5p (miR-515-5p) on the cisplatin sensitivity of non-small cell lung cancer (NSCLC).

Methods

The expression of miR-515-5p in BEAS-2B, A549 and A549/DDP cells was detected by real-time fluorescent quantitative PCR (RT-qPCR). A549/DDP cells were divided into NC group, miR-515-5p group, si-RING1 group and miR-515-5p inh+ si-RING1 group. CCK-8 assay was used to assess the proliferation of A549/DDP cells. WB was performed to measure the expression of RING1 and apoptosis related proteins. Dual luciferase reporter gene system was used to verify the targeting relationship between miR-515-5p and RING1.

Results

The expression of miR-515-5p in BEAS-2B, A549 and A549/DDP cells were (1.00±0.03), (0.61±0.03), (0.31±0.04), respectively. A549/DDP cells were transfected with miR-515-5p mimics, the OD₄₅₀ values of cells in NC group and miR-515-5p group at 24, 48, 72 and 96 h were (0.61±0.02), (0.52±0.02), (0.84±0.02), (0.68±0.04), (1.03±0.03), (0.79±0.04), (1.08±0.03), (0.86±0.04), respectively. The differences of those items were all statistically significant (all P<0.05). The expression of cleaved caspase-3, cleaved PARP and Bax in miR-515-5p group were significantly higher than those in NC group, and the expression of Bcl-2 were significantly lower than those in NC group. A549/DDP cells were transfected with si-RING1 and miR-515-5p inhibitor, the OD₄₅₀ values of cells in NC group, si-RING1 group and miR-515-5p inh+ si-RING1 group at 24, 48, 72 and 96 h were (0.63±0.02), (0.49±0.04), (0.58±0.03), (0.83±0.03), (0.62±0.05), (0.80±0.02), (1.05±0.04), (0.76±0.03), (1.02±0.04), (1.15±0.05), (0.86±0.03), (1.12±0.06), respectively. The differences of those items were all statistically significant (all P<0.05). The expression of cleaved caspase-3, cleaved PARP and Bax in si-RING1 group were significantly higher than those in NC group and miR-515-5p inh+ siRING1 group, and the expression of Bcl-2 were significantly lower than those in NC group and miR-515-5p inh+ siRING1 group.

Conclusion

miR-515-5p enhanced the cisplatin sensitivity of A549/DDP cells through targeting downregulate the expression of RING1.

Key words: MicroRNA-515-5p, Ring finger protein 1, Non-small cell lung cancer, Cisplatin sensitivity

Rui Xu, Yong Qian, Haiyan Zhang. Study on the mechanism of miR-515-5p enhanced the cisplatin sensitivity of NSCLC A549/DDP cells via targeting RING1[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2021, 14(05): 559-563.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhfbjbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-6902.2021.05.003

https://zhfbjbzz.cma-cmc.com.cn/EN/Y2021/V14/I05/559

Figures/Tables 5

References 31

1	Yang B, Zheng D, Zeng U, et al. Circulating tumor cells predict prognosis following secondline AZD 9291 treatment in EGFR-T790M mutant non-small cell lung cancer patients[J]. J BUON, 2018, 23(4): 1077-1081.
2	Galluzzi L, Senovilla L, Vitale I, et al. Molecular mechanisms of cisplatin resistance[J]. Oncogene, 2012, 31(15): 1869-1883.
3	肖丹，杜琴，贺斌峰，等. YAP调控肺癌A549/DDP细胞顺铂耐药性的机制分析[J/CD]. 中华肺部疾病杂志(电子版), 2019, 12(6): 697-701.
4	Rao C, Miao X, Zhao G, et al. MiR-219a-5p enhances cisplatin sensitivity of human non-small cell lung cancer by targeting FGF9[J]. Biomed Pharmacoth, 2019, 114: 108662.
5	Zhou H, Li J, Zhang Z, et al. RING1 and YY1 binding protein suppresses breast cancer growth and metastasis[J]. Int J Oncol, 2016, 49(6): 2442-2452.
6	Zhou Y, Wan C, Liu Y, et al. Polycomb group oncogene RING1 is over-expressed in non-small cell lung cancer[J]. Pathol Oncol Res, 2014, 20(3): 549-556.
7	Shen J, Li P, Shao X, et al. The E3 ligase RING1 targets p53 for degradation and promotes cancer cell proliferation and survival[J]. Cancer Res, 2018, 78(2): 359-371.
8	Xiong Y, Hu B, Wei L, et al. Upregulated expression of polycomb protein Ring1 contributes to poor prognosis and accelerated proliferation in human hepatocellular carcinoma[J]. Tumour Biol, 2015, 36(12): 9579-9588.
9	Hu B, Zhang H, Wang Z, et al. LncRNA CCAT1/miR-130a-3p axis increases cisplatin resistance in non-small-cell lung cancer cell line by targeting SOX4[J]. Cancer Biol Ther, 2017, 18(12): 974-983.
10	Xu G, Zhang Z, Zhang L, et al. miR-4326 promotes lung cancer cell proliferation through targeting tumor suppressor APC2[J]. Molecul Cellul Biochem, 2018, 443(1-2): 151-157.
11	Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method[J]. Methods, 2001, 25(4): 402-408.
12	Chen K, Abuduwufuer A, Zhang H, et al. SNHG7 mediates cisplatin-resistance in non-small cell lung cancer by activating PI3K/AKT pathway[J]. Eur Rev Med Pharmacol Sci, 2019, 23(16): 6935-6943.
13	Guo J, Jin D, Wu Y, et al. The miR 495-UBE2C-ABCG2/ERCC1 axis reverses cisplatin resistance by downregulating drug resistance genes in cisplatin-resistant non-small cell lung cancer cells[J]. EBioMedicine, 2018, 35: 204-221.
14	Li J H, Liu S, Zhou H, et al. starBase v2.0: decoding miRNA-ceRNA, miRNA-ncRNA and protein-RNA interaction networks from large-scale CLIP-Seq data[J]. Nucleic Acids Res, 2014, 42(Database issue): D92-D97.
15	Du L, Pertsemlidis A. microRNA regulation of cell viability and drug sensitivity in lung cancer[J]. Expert Opin Biol Ther, 2012, 12(9): 1221-1239.
16	Wang S, Li MY, Liu Y, et al. The role of microRNA in cisplatin resistance or sensitivity[J]. Expert Opin Ther Targets, 2020, 24(9): 885-897.
17	Lu X, Liu R, Wang M, et al. MicroRNA-140 impedes DNA repair by targeting FEN1 and enhances chemotherapeutic response in breast cancer[J]. Oncogene, 2019, 39(1): 234-247.
18	Rong F, Liu L, Zou C, et al. MALAT1 promotes cell tumorigenicity through regulating miR-515-5p/EEF2 axis in non-small cell lung cancer[J]. Cancer Manag Res, 2020, 12: 7691-7701.
19	Liu J, Eckert MA, Harada BT, et al. m(6)A mRNA methylation regulates AKT activity to promote the proliferation and tumorigenicity of endometrial cancer[J]. Nature Cell Biol, 2018, 20(9): 1074-1083.
20	Van Leenders GJ, Dukers D, Hessels D, et al. Polycomb-group oncogenes EZH2, BMI1, and RING1 are overexpressed in prostate cancer with adverse pathologic and clinical features[J]. Eur Urol, 2007, 52(2): 455-463.
21	Yuan XW, Yan TQ, Tong H. Effect of miR-515-5p on proliferation and drug sensitivity of retinoblastoma cells[J]. Cancer Manag Res, 2020, 12: 12087-12098.
22	Zhong Y, Wang D, Ding Y, et al. Circular RNA circ_0032821 contributes to oxaliplatin (OXA) resistance of gastric cancer cells by regulating SOX9 via miR-515-5p[J]. Biotechnol Lett, 2021, 43(2): 339-351.
23	Han Y, Li F, Xie J, et al. PVT1 mediates cell proliferation, apoptosis and radioresistance in nasopharyngeal carcinoma through regulating miR-515-5p/PIK3CA axis[J]. Cancer Manag Res, 2020, 12: 10077-10090.
24	Qiao K, Ning S, Wan L, et al. LINC00673 is activated by YY1 and promotes the proliferation of breast cancer cells via the miR-515-5p/MARK4/Hippo signaling pathway[J]. J Exp Clin Cancer Res, 2019, 38(1): 418.
25	Wang D, Liu K, Chen E. LINC00511 promotes proliferation and invasion by sponging miR-515-5p in gastric cancer[J]. Cell Molecul Biol Lett, 2020, 25: 4.
26	Zhang Y, Shi Z, Li Z, et al. Circ_0057553/miR-515-5p regulates prostate cancer cell proliferation, apoptosis, migration, invasion and aerobic glycolysis by targeting YES1[J]. Onco Targets Ther, 2020, 13: 11289-11299.
27	Zhang X, Zhou J, Xue D, et al. MiR-515-5p acts as a tumor suppressor via targeting TRIP13 in prostate cancer[J]. Int J Biol Macromol, 2019, 129: 227-232.
28	Ni JS, Zheng H, Ou YL, et al. miR-515-5p suppresses HCC migration and invasion via targeting IL6/JAK/STAT3 pathway[J]. Surg Oncol, 2020, 34: 113-120.
29	Guo F, Li S, Guo C, et al. Circular RNA circMAGI3 accelerates the glycolysis of non-small cell lung cancer through miR-515-5p/HDGF[J]. Am J Transl Res, 2020, 12(7): 3953-3963.
30	Wang Y, Zhang Q. Long noncoding RNA MALAT1 knockdown inhibits proliferation, migration, and invasion and promotes apoptosis in non-small-cell lung cancer cells through regulating miR-515-3p/TRIM65 axis[J]. Cancer Biother Radiopharm, 2020, doi: 10.1089/cbr.2020.3730.
31	Li J, Tang Z, Wang H, et al. CXCL6 promotes non-small cell lung cancer cell survival and metastasis via down-regulation of miR-515-5p[J]. Biomed Pharmacother, 2018, 97: 182-1188.

分　组	24 h	48 h	72 h	96 h
NC组	0.61±0.02	0.84±0.02	1.03±0.03	1.08±0.03
miR-515-5p组	0.52±0.02^a	0.68±0.04^a	0.79±0.04^a	0.86±0.04^a

分　组	24 h	48 h	72 h	96 h
NC组	0.61±0.02	0.84±0.02	1.03±0.03	1.08±0.03
miR-515-5p组	0.52±0.02^a	0.68±0.04^a	0.79±0.04^a	0.86±0.04^a

分　组	24 h	48 h	72 h	96 h
NC组	0.63±0.02	0.83±0.03	1.05±0.04	1.15±0.05
si-SING1组	0.49±0.04^a	0.62±0.05^a	0.76±0.03^a	0.86±0.03^a
miR-515-5p inh　+si-RING1组	0.58±0.03^b	0.80±0.02^b	1.02±0.04^b	1.12±0.06^b

分　组	24 h	48 h	72 h	96 h
NC组	0.63±0.02	0.83±0.03	1.05±0.04	1.15±0.05
si-SING1组	0.49±0.04^a	0.62±0.05^a	0.76±0.03^a	0.86±0.03^a
miR-515-5p inh　+si-RING1组	0.58±0.03^b	0.80±0.02^b	1.02±0.04^b	1.12±0.06^b

[1]	Lisi Liang, Jie Li, Shuai He, Yanjun Lai, Ming Liu, Lin Zhang. Significance of MMP-9， MMP-2， TLR4， HE4 in early diagnosis of non-small cell lung cancer [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(05): 756-761.
[2]	Miao Lai, Xin Jing, Guizhen Li, Yi Li. Clinicopathological and prognostic significance of EGFR mutant subtypes in patients with non-small cell lung cancer [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(05): 731-737.
[3]	Hui Yang, Lijuan Guo, Xiaodan Feng, Jing LI, Chengmo Huang, Xingrui Cai, Yingjiao Qin, Yuanli Wang. Expression characteristics and prediction analysis of platinum-resistant mi RNA in non-small cell lung cancer [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(05): 719-724.
[4]	Mengmeng Zhao, Jie Huang, Ronghuan Yu, Baoqing Wang. Overexpression of the small GTPase Rab32 inhibits the invasive growth of non-small cell lung cancer cells [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(04): 512-518.
[5]	Guiping Zhang, Yonglin Qiu, Qiting Zhan, Ledong Sun. Prognostic significance of serum Ape1/Ref-1 for the radioactive lung injury in patients with advanced non-small cell lung cancer [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(04): 519-523.
[6]	Xiaoyu Han, Keyu Li, Zhifei Zhao, Jianping Gao. Prognostic significance of SNHG17 overexpression for non-small cell lung cancer resection [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(04): 543-547.
[7]	Song Liu, Jinzhao Zhang, Yanyun Jia. Relationship between reduced response to pabolizumab in treatment of advanced non-small cell lung cancer and antibiotic preconditioning [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(04): 553-557.
[8]	Duo Li, Zhaozhao Hao, Yanwei Chen, Yandong Nan. Correlation analysis of serum PTX3 expression and bone metastasis in non-small cell lung cancer [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(04): 558-562.
[9]	Lili Bai, Yu Jiang, Liangliang Huang, Ying BAI, Min Zhang. Application of occupational therapy in postoperative rehabilitation of patients with non-small cell lung cancer [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(03): 411-415.
[10]	Wei Cui, Yi Deng, Suyi Ye, Jing Li, Xiaoming Chen, Jing Zhang, Rongde Xu. Clinical efficacy and safety of drug-eluting bead bronchial artery chemoembolization in the treatment of rare non-small cell lung cancer [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2024, 12(04): 303-310.
[11]	Wei Cui, Suyi Ye, Yi Deng, Xiaoming Chen, Jing Zhang, Jing Li, Rongde Xu. Clinical efficacy and safety analysis of bronchial artery chemoembolization with drug-loaded microspheres in the treatment of refractory non-small cell lung cancer [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2024, 12(04): 311-316.
[12]	Wei Cui, Jing Li, Xiaoming Chen, Jing Zhang, Yi Deng, Rongde Xu. Research progress of drug-eluting beads bronchial arterial chemoembolization in the treatment of nonsmall cell lung cancer [J]. Chinese Journal of Interventional Radiology(Electronic Edition), 2024, 12(04): 289-295.
[13]	Jianqiao Cai, Lei Jiang. Comparison of uniportal video-assisted thoracoscopic and thoracotomy double sleeve lobectomy for the treatment of non-small cell lung cancer [J]. Chinese Journal of Thoracic Surgery(Electronic Edition), 2024, 11(04): 225-230.
[14]	Zijian Li, Rui Wang, Yunpeng Zhong, Dixuan Zhang, Yunjuan Liang, Chao Yang, Jianxing He, Shuben Li. Clinical application of lung autotransplantation in thoracic malignancies [J]. Chinese Journal of Thoracic Surgery(Electronic Edition), 2024, 11(03): 193-200.
[15]	Xunfu Zhang, Jinshan Ma, Yunlong Jiang, Tuoleheng Jianati, Changjian Hou, Silajiding Sawuti. Expression and clinicopathological significance of GATA3 protein in non-small cell lung cancer tissues [J]. Chinese Journal of Thoracic Surgery(Electronic Edition), 2024, 11(03): 175-179.

Please choose a citation manager

Content to export