Inhibitory of miR-106a-5p regulating PTEN on the proliferation and migration of nasopharyngeal carcinoma cells SUNE2

doi:10.3877/cma.j.issn.2095-1221.2020.04.004

Abstract

Abstract:

Objective

To investigate the effect of miR-106a-5p on the proliferation and migration of nasopharyngeal carcinoma cells SUNE2.

Methods

The nasopharyngeal carcinoma cells SUNE2 cultured in vitro were divided into Control group (without treatment), Anti-NC group (transfected with inhibitor control) and Anti-miR-106a-5p group (transfected with miR-106a-5p inhibitor), as well as Anti-miR-106a-5p+si-NC group (transfected with miR-106a-5p inhibitor and siRNA control) and Anti-miR-106a-5p+si-PTEN group (transfected with miR-106a-5p inhibitor and PTEN siRNA). Realtime PCR was used to determine the expression of miR-106a-5p, MTT and Transwell were used to detect proliferation rate, invasion and migration ability of cells, respectively. The expression levels of vimentin and E-cadherin protein were detected by Western blot. The online target gene prediction software suggests that the target gene of miR-106a-5p may be PTEN, and the dual luciferase reporter system examines the targeting relationship between miR-106a-5p and PTEN. The independent sample t-test was used for the comparison between two groups, the ANOVA analysis was used for the multiple group comparison, and the LSD-t test was used for the pairwise comparison of the components.

Results

Compared with NP69 in normal nasopharyngeal epithelial cells, the expression level of miR-106a-5p in nasopharyngeal carcinoma cells CNE-2, HK1, and SUNE2 were increased (1.00±0.11 vs 1.84±0.13, 2.19±0.14, 2.87±0.25), the difference was statistically significant (P < 0.05) .Compared with the Control and Anti-NC group, the expression level of miR-106a-5p (1.00±0.10, 0.99±0.12 vs 0.76±0.08), the OD values (0.56±0.05, 0.57±0.04 vs 0.32±0.02), cell invasion (128.47±11.65, 129.84±10.93 vs 85.12±6.75), the number of migrations (182.51±14.81, 180.68±17.64 vs 122.01±11.62) as well as the expression level of vimentin protein (0.84±0.09, 0.82±0.07 vs 0.30±0.05) of nasopharyngeal cancer cells in the Anti-miR-106a-5p group were significantly reduced. While the expression level of E-cadherin protein was increased (0.29±0.04, 0.28±0.05 vs 0.76±0.08) (P < 0.05). Compared with the Anti-miR-106a-inhibitor+si-NC group, in the Anti-miR-106a-inhibitor+si-PTEN group, the OD value (0.33±0.03 vs 0.52±0.05), the number of invasions (84.16±5.91 vs 105.79±8.63) and migrations (118.42±10.25 vs 164.28±12.05) as well as the expression of vimentin protein (0.34±0.06 vs 0.68±0.05) were all significantly increased. While the expression of E-cadherin protein was decreased (0.72±0.06 vs 0.29±0.05) (P < 0.05) .

Conclusion

miR-106a-5p could inhibit the proliferation and migration of nasopharyngeal carcinoma cells SUNE2 by targeted regulation of PTEN.

Key words: Nasopharyngeal carcinoma, MiR-106a-5p, PTEN, Invasion, Proliferation

Zhuan Li, Lihua Rao, Hong Gao, Kuirong Wang. Inhibitory of miR-106a-5p regulating PTEN on the proliferation and migration of nasopharyngeal carcinoma cells SUNE2[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2020, 10(04): 219-225.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2020.04.004

https://zhxbygxbzz.cma-cmc.com.cn/EN/Y2020/V10/I04/219

Figures/Tables 12

References 24

1	Lu S, Wei J, Sun F, et al. Late sequelae of childhood and adolescent nasopharyngeal carcinoma survivors after radiation therapy[J]. Int J Radia Oncol Biol Phys, 2019, 103(1):45-51.
2	Tu C, Zeng Z, Qi P, et al. Identification of genomic alterations in nasopharyngeal carcinoma and nasopharyngeal carcinoma-derived Epstein-Barr virus by whole-genome sequencing[J]. Carcinogenesis, 2018, 39(12): 1517-1528.
3	Müller S, Bley N, Glaß M, et al. IGF2BP1 enhances an aggressive tumor cell phenotype by impairing miRNA-directed downregulation of oncogenic factors[J]. Nucleic Acids Res, 2018, 46(12):6285-6303.
4	Guo S, Lin WN, Hu Y, et al. Ultrahigh-throughput droplet microfluidic device for single-cell miRNA detection with isothermal amplification[J]. Lab Chip, 2018, 18(13):1914-1920.
5	Adams BD, Arem H, Hubal MJ, et al. Exercise and weight loss interventions and miRNA expression in women with breast cancer[J]. Breast Cancer Research Treat, 2018, 170(1):55-67.
6	Luo B, Kang N, Chen Y, et al. Oncogene miR-106a promotes proliferation and metastasis of prostate cancer cells by directly targeting PTEN in vivo and in vitro[J]. Minerva Med, 2018, 109(1):24-30.
7	Hu B, Cai H, Zheng R, et al. Long non-coding RNA 657 suppresses hepatocellular carcinoma cell growth by acting as a molecular sponge of miR-106a-5p to regulate PTEN expression[J]. Int J Biochem Cell Biol, 2017, 92(11):34-42.
8	Bian X, Gao J, Luo F, et al. PTEN deficiency sensitizes endometrioid endometrial cancer to compound PARP-PI3K inhibition but not PARP inhibition as monotherapy[J]. Oncogene, 2018, 37(3):341-351.
9	Lu J, Mu X, Yin Q, et al. miR-106a contributes to prostate carcinoma progression through PTEN[J]. Oncol Lett, 2019, 17(1): 1327-1332.
10	Liu J, Huang Y, Wang H, et al. MiR-106a-5p promotes 5-FU resistance and the metastasis of colorectal cancer by targeting TGFβR2[J]. Int J Clin Exp Pathol, 2018, 11(12):5622-5634.
11	丁莎莎，陈晓丽，陈子兴, 等. MicroRNA-106a在非小细胞肺癌中的表达及意义[J]. 现代肿瘤医学, 2015, 23(16):2289-2291.
12	Shi B, Ma C, Liu G, et al. MiR-106a directly targets LIMK1 to inhibit proliferation and EMT of oral carcinoma cells[J]. Cell Mol Biol Lett, 2019, 24(1):1-13.
13	Zare M, Bastami M, Solali S, et al. Aberrant miRNA promoter methylation and EMT-involving miRNAs in breast cancer metastasis: diagnosis and therapeutic implications[J]. J Cell Physiol, 2018, 233(5):3729-3744.
14	Lamprecht S, Kaller M, Schmidt EM, et al. PBX3 is part of an EMT regulatory network and indicates poor outcome in colorectal cancer[J]. Clin Cancer Res, 2018, 24(8):1974-1986.
15	Zhang Q, Li X, Li X, et al. LncRNA H19 promotes epithelial-mesenchymal transition (EMT) by targeting miR-484 in human lung cancer cells[J]. J Cell Biochem, 2018, 119(6):4447-4457.
16	Zhou Z, Zhang HS, Liu Y, et al. Loss of TET1 facilitates DLD1 colon cancer cell migration via H3K27me3-mediated down-regulation of E-cadherin[J]. J Cell Physiol, 2018, 233(2):1359-1369.
17	Dufresne S, Rébillard A, Muti P, et al. A review of physical activity and circulating miRNA expression: implications in cancer risk and progression[J]. Cancer Epidemiol Biomarkers Prev, 2018, 27(1):11-24.
18	Lo UG, Pong RC, Yang D, et al. IFNγ-induced IFIT5 promotes epithelial-to-mesenchymal transition in prostate cancer via miRNA processing[J]. Cancer Res, 2019, 79(6):1098-1112.
19	Ding M, Van der Kwast TH, Vellanki RN, et al. The mTOR Targets 4E-BP1/2 restrain tumor growth and promote hypoxia tolerance in PTEN-driven prostate cancer[J]. Mol Cancer Res, 2018, 16(4):682-695.
20	Kim SM, Nguyen TT, Ravi A, et al. PTEN deficiency and AMPK activation promote nutrient scavenging and anabolism in prostate cancer cells[J]. Cancer Discov, 2018, 8(7):866-883.
21	He F, Ma N, Midorikawa K, et al. Taurine exhibits an apoptosis-inducing effect on human nasopharyngeal carcinoma cells through PTEN/Akt pathways in vitro[J]. Amino Acids, 2018, 50(12):1749-1758.
22	Hsu YL, Hung JY, Chang WA, et al. Hypoxic lung-Cancer-derived extracellular vesicle MicroRNA-103a increases the oncogenic effects of macrophages by targeting PTEN[J]. Mol Ther, 2018, 26(2):568-581.
23	Lopez C, Abuel-Haija M, Pena L, et al. Novel germline PTEN mutation associated with cowden syndrome and osteosarcoma[J]. Cancer Genomics Proteomics, 2018, 15(2):115-120.
24	Yang Y, Guo JX, Shao ZQ. miR-21 targets and inhibits tumor suppressor gene PTEN to promote prostate cancer cell proliferation and invasion: An experimental study[J]. Asian Pac J Trop Med, 2017, 10(1):87-91.

基因	序列（5'-3'）	预期扩增产物长度（bp）
miR-106a-5p	上游ATCCAGTGCTGTCGTG	92
?	下游TGCTAAAAGTGCTTACAGTG	?
U6（内参）	上游TGCGGGTGCTCGCTTCGGCAGC	96
?	下游GTGCAGGGTCCGAGGT	?

基因	序列（5'-3'）	预期扩增产物长度（bp）
miR-106a-5p	上游ATCCAGTGCTGTCGTG	92
?	下游TGCTAAAAGTGCTTACAGTG	?
U6（内参）	上游TGCGGGTGCTCGCTTCGGCAGC	96
?	下游GTGCAGGGTCCGAGGT	?

分组	miR-106a-5p
对照组	1.00±0.10
Anti-NC组	0.99±0.12
Anti-miR-106a-5p组	0.31±0.05^ab
F值	157.015
P值	< 0.001

分组	miR-106a-5p
对照组	1.00±0.10
Anti-NC组	0.99±0.12
Anti-miR-106a-5p组	0.31±0.05^ab
F值	157.015
P值	< 0.001

分组	OD值	侵袭数目（个）	迁移数目（个）	vimentin	E-cadherin
对照组	0.56±0.05	128.47±11.65	182.51±14.81	0.84±0.09	0.29±0.04
Anti-NC组	0.57±0.04	129.84±10.93	180.68±17.64	0.82±0.07	0.28±0.05
Anti-miR-106a-5p组	0.32±0.02^ab	85.12±6.75^ab	122.01±11.62^ab	0.30±0.05^ab	0.76±0.08^ab
F值	120.200	58.070	48.046	163.277	193.457
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

Please choose a citation manager

Content to export