Mechanism of miR-214 on the proliferation and apoptosis of oropharynx squamous cell carcinoma cell line by regulating Notch1 signal

doi:10.3877/cma.j.issn.2095-1221.2019.06.005

Abstract

Abstract:

Objective

To study the mechanism of miR-214 on the proliferation, invasion and apoptosis of oropharyngeal squamous cell carcinoma (OSCC) cell line by regulating Notch 1 signal.

Methods

OSCC cell lines were purchased from ATCC cell bank and divided into three groups: inhibition group (IN group) , simulation group (SI group) and control group (CO group) . Notch 1 protein, mRNA expression, apoptosis, migration and proliferation were analyzed by Western blot, qRT-PCR, TUNEL, invasion experiment and CCK-8. One way ANOVA was used to compare differences among the three groups, LSD-t test was used for comparison between two groups.

Result

The expression of mir-214m RNA in the cells of IN group, SI group and CO group were (1.15 ±0.26) , (3.34±0.89) , (2.08±0.67) respectively, and there was significant difference between the SI group and IN group, SI group and CO group (t = 5.281, P = 0.002; t = 2.529, P = 0.045) ; The apoptosis rates of OSCC cells in the three groups were (0.78±0.10) ﹪, (0.32±0.06) ﹪, (0.56±0.12) ﹪, respectively. The difference between IN group and CO group, IN group and SI group were statistically significant (t = 3.149, P = 0.020; t = 8.823, P < 0.001) ; The proliferation rates of cancer cells in the three groups were: (0.65±0.05) ﹪, (1.26±0.06) ﹪, (1.89±0.04) ﹪, respectively. The difference between IN group and SI group, IN group and CO group were statistically significant (t = 11.056, P < 0.001; t = 27.394, P < 0.001) . The difference between CO group and SI group was statistically significant (t = 12.365, P < 0.001) ; the invasive rates of cancer cells in the three groups were: (0.13±0.02) ﹪, (0.89±0.13) ﹪, (0.48±0.11) ﹪, respectively. The difference between SI group and CO group, SI group and IN group were tatistically significant (t = 4.176, P = 0.006; t = 10.147, P < 0.001) ; The levels of Notch1 protein in the three groups were (2.38±1.34) , (0.94 ± 0.23) , (1.76 ± 0.67) , respectively. There was a significant difference between SI group and CO group (t = 2.588, P = 0.041; t = 2.368, P = 0.056) ; The expression of Notch1 mRNA in the three groups were (4.26 ± 1.06) , (1.45 ± 0.38) , (2.46 ± 0.87) , respectively. The difference was statistically significant (t = 2.935, P = 0.026; t = 5.583, P = 0.001) .

Conclusion

miR-214 levels are elevated and may be inversely related to the proliferation of OSCC cell lines. MiR-214 inhibits Notch1 signaling pathway expression, promotes growth and invasion of cancer cells, and inhibites apoptosis of cancer cells.

Key words: miR-214, Notch1, Oropharyngeal squamous cell carcinoma

Shaobin Liu, Zichen Liu, Changjuan Meng, Cuili Zhang. Mechanism of miR-214 on the proliferation and apoptosis of oropharynx squamous cell carcinoma cell line by regulating Notch1 signal[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2019, 09(06): 345-350.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhxbygxbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-1221.2019.06.005

https://zhxbygxbzz.cma-cmc.com.cn/EN/Y2019/V09/I06/345

Figures/Tables 8

References 20

1	陈睿，杨朝晖，陈伟良, 等. 18例侵犯颈动脉的复发性口腔、口咽鳞状细胞癌的挽救性手术治疗效果评价[J]. 中国口腔颌面外科杂志, 2017, 15(3):277-281.
2	Pai SI, Faivre S, Licitra L, et al. Comparative analysis of the phase III clinical trials of anti-PD1 monotherapy in head and neck squamous cell carcinoma patients (CheckMate 141 and KEYNOTE 040)[J]. J Immunother Cancer, 2019, 7(1):96-101.
3	Oguejiofor K, Galletta-Williams H, Dovedi SJ, et al. Distinct patterns of infiltrating CD8⁺ T cells in HPV⁺ and CD68 macrophages in HPV^-oropharyngeal squamous cell carcinomas are associated with better clinical outcome but PD-L1 expression is not prognostic[J]. Oncotarget, 2017, 8(9):14416-14427.
4	林璐，韩影，熊浩峰, 等. 口腔鳞状细胞癌复发的临床和病理因素分析[J]. 北京医学, 2018, 40(10):47-51.
5	Molony P, Kharytaniuk N, Boyle S, et al. Impact of positive margins on outcomes of oropharyngeal squamous cell carcinoma according to p16 status[J]. Head Neck, 2017, 39(8):1680-1688.
6	Jouhi L, Atula T, Saarilahti K, et al. Changing trends in the management of the neck in oropharyngeal squamous cell carcinoma[J]. Head Neck, 2017, 39(7):1412-1420.
7	Hu JL, He GY, Lan XL, et al. Inhibition of ATG12-mediated autophagy by miR-214 enhances radiosensitivity in colorectal cancer[J]. Oncogenesis, 2018, 7(2):16-18.
8	Li F, Ma K, Sun MH, et al. Identification of the tumor-suppressive function of circular RNA ITCH in glioma cells through sponging miR- 214 and promoting linear ITCH expression[J]. Am J Transl Res, 2018, 10(5):1373-1386.
9	Wang XY, Zhang HY, Bai M, et al. Exosomes serve as nanoparticles to deliver Anti-miR-214 to reverse chemoresistance to cisplatin in gastric cancer[J]. Molecular Therapy, 2018, 26(3):774-783.
10	Xu LA, Gu LA, Tao XF, et al. Effect of dioscin on promoting liver regeneration via activating Notch1/Jagged1 signal pathway[J]. Phytomedicine, 2018, 38(13):107-117.
11	Dickinson A, Xu M, Silén S, et al. Newly detected DNA viruses in juvenile nasopharyngeal angiofibroma (JNA) and oral and oropharyngeal squamous cell carcinoma (OSCC/OPSCC)[J]. Am J Transl Res, 2019, 276(2):613-617.
12	Hai L, Zhang C, Li T, et al. Notch1 is a prognostic factor that is distinctly activated in the classical and proneural subtype of glioblastoma and that promotes glioma cell survival via the NF- κB(p65) pathway[J]. Cell Death Dis, 2018, 9(2):158-164.
13	Lilja AM, Rodilla V, Huyghe M, et al. Clonal analysis of Notch1-expressing cells reveals the existence of unipotent stem cells that retain long-term plasticity in the embryonic mammary gland[J]. Nat Cell Biol, 2018, 20(6):677-680.
14	赵泽亮. 口咽和非口咽鳞状细胞癌中Trk蛋白的表达:人乳头瘤病毒感染的临床意义和可能的作用[J]. 中国口腔颌面外科杂志, 2018, 16(6):35.
15	王晓乐，贾荣. 口腔鳞状细胞癌患者年龄分布与肿瘤分子标记物的关系研究[J]. 口腔医学研究, 2019, 35(3):238-241.
16	张海东，龚单春，刘亚群, 等. 循环肿瘤细胞在头颈部鳞状细胞癌中意义的初步研究[J]. 中华耳鼻咽喉头颈外科杂志, 2018, 53(1):39-44.
17	Corbett J, Wilke D, Trites J, et al. Orbital mass as first presentation of metastatic p16-positive oropharyngeal squamous cell carcinoma[J]. Curr Oncol, 2017, 24(6):e551-e554.
18	Rubek N, Channir HI, Charabi BW, et al. Primary transoral robotic surgery with concurrent neck dissection for early stage oropharyngeal squamous cell carcinoma implemented at a Danish head and neck cancer center: a phase II trial on feasibility and tumour margin status[J]. Eur Arch Otorhinolaryngol, 2017, 274(5):2229-2237.
19	Wang F, Tan WH, Liu W, et al. Effects of miR-214 on cervical cancer cell proliferation, apoptosis and invasion via modulating PI3K/AKT/mTOR signal pathway[J].Eur Rev Med Pharmacol Sci, 2018, 22(7):1891-1898.
20	Guanen Q, Junjie S, Baolin W, et al. MiR-214 promotes cell meastasis and inhibites apoptosis of esophageal squamous cell carcinoma via PI3K/AKT/mTOR signaling pathway[J]. Biomed Pharmacother, 2018, 105(9):350-361.

蛋白	基因	引物序列
Notch1	F	5'-GACATCACGGATCATATGGA-3'
?	R	5'-CTCGCATTGACCATTCAAAC-3'
miR-214	F	5'-TGCGGTAGCAGCACATAATGG-3'
?	R	5'-CCAGTGCAGGGTCCGAGGT-3'
GAPDH	F	5'-TGAACGGGAAGCTCACTGG-3'
?	R	5'-TCCACCACCCTGTTGCTGTA-3'

蛋白	基因	引物序列
Notch1	F	5'-GACATCACGGATCATATGGA-3'
?	R	5'-CTCGCATTGACCATTCAAAC-3'
miR-214	F	5'-TGCGGTAGCAGCACATAATGG-3'
?	R	5'-CCAGTGCAGGGTCCGAGGT-3'
GAPDH	F	5'-TGAACGGGAAGCTCACTGG-3'
?	R	5'-TCCACCACCCTGTTGCTGTA-3'

分组	实验次数	细胞凋亡率	增殖率	侵袭率
CO组	3	0.56±0.12	1.26±0.06	0.48±0.11
IN组	3	0.78±0.10^a	0.65±0.05^a	0.13±0.02^a
SI组	3	0.32±0.06^b	1.89±0.04^b	0.89±0.13^b
F值	?	17.013	299.627	48.369
P值	?	0.003	< 0.001	< 0.001

分组	实验次数	细胞凋亡率	增殖率	侵袭率
CO组	3	0.56±0.12	1.26±0.06	0.48±0.11
IN组	3	0.78±0.10^a	0.65±0.05^a	0.13±0.02^a
SI组	3	0.32±0.06^b	1.89±0.04^b	0.89±0.13^b
F值	?	17.013	299.627	48.369
P值	?	0.003	< 0.001	< 0.001

[1]	Han Xie, Qiong Chen, Yichun Wang. Inhibitory effect of endostatin on pulmonary fibrosis in mice through Notch1/platelet-derived growth factor signaling pathway [J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2019, 12(06): 361-366.
[2]	Ke Zhang, Zhiqiang Wu, Wenwei Cai, Shibing Wang. Protective mechanism of total flavonoids from Chrysanthemum morifolium in mice with cardiac hypertrophy based on Notch1 signaling pathway [J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2019, 12(05): 295-300.
[3]	Lina Zhao, Jinghua Wu, Guocheng Liu, Ruixia Ma. Clinical significance of expressions of Notch1 protein and PPAR-γ protein in placenta of patients with early onset severe preeclampsia [J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2019, 15(02): 164-170.
[4]	Ziyun Shi, Yanchuan Li, Feifei Liu, Ya Zhang, Xiaohua Yuan, Shue Wang. Effects of the activation of Notch1 signaling pathway in hypoxia-induced angiogenesis of human umbilical vein endothelial cells [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2020, 10(03): 163-171.
[5]	Wei Zheng, Hulin Chang, Jun Hai, Xiaoxue Song, Lixue Du. Effect of Ang Ⅱ on the activation and proliferation of hepatic stellate cell LX2 by activating the Notch1/Sox2 pathway [J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2018, 08(03): 156-160.

Please choose a citation manager

Content to export