miR-191-5p通过靶向TJP1调控Cal-27细胞增殖、侵袭和迁移的体外研究

doi:10.13591/j.cnki.kqyx.2023.11.004

Abstract

Abstract:

Objective To investigate the biological function and mechanism of miR-191-5p regulating OSCC cells. Methods The expression of miR-191-5p in oral squamous cell carcinoma patients was analyzed by bioinformatics method, and the possible target gene TJP1 was screened out through the relevant database. The targeting relationship between the two was verified by dual-luciferase experiment. In vitro transfection was divided into blank, inhibitor NC (negative control group) and miR-191-5p inhibitor (knockdown miR-191-5p group). After knocking down miR-191-5p, the transfection efficiency was verified by qRT-PCR, and the effects of miR-191-5p on the proliferation, invasion and migration ability of Cal-27 cells were detected by CCK-8 proliferation experiment, cell cloning experiment, Transwell and scratch experiment. Western blotting detected the expression of TJP1 protein and EMT-associated proteins. siRNA-TJP1 was transfected into miR-191-5p low-expressing cells and the effect of simultaneous down-regulation of TJP1 on the proliferation and migration capacity of Cal-27 cells was observed. Results miR-191-5p expression was up-regulated in OSCC tissues and cells, while down-regulation of miR-191-5p inhibited the proliferation, invasion and migration capacity of Cal-27 cells and EMT tendency. miR-191-5p targeted and inhibited the expression of TJP1, while down-regulation of TJP1 reversed the inhibition of Cal-27 cell proliferation and migration by knocking down miR-191-5p. Conclusion The proliferation, invasion and migration of OSCC cells is inhibited by targeting TJP1 under miR-191-5p, and miR-191-5p may increase tumor metastasis by regulating the EMT pathway.

Key words: oral squamous cell carcinoma, miR-191-5p, tight junction protein 1(TJP1), proliferation, invasion and migration, epithelial-mesenchymal transition

CLC Number:

R739.85

TONG Xuexi, ZHAO Gang. In vitro study on miR-191-5p targeting TJP1 to regulate the proliferation, invasion and migration of Cal-27 cells[J]. Stomatology, 2023, 43(11): 981-988.

Figures/Tables 11

Tab.1

Fig.1

Fig.2

Fig.3

Fig.4

Fig.5

Fig.6

Fig.7

Fig.8

Fig.9

Fig.10

References 21

[1]	Peng Y, Croce CM. The role of micro RNAs in human cancer[J]. Signal Transduct Target Ther, 2016, 1:15004.
[2]	Hayes J, Peruzzi PP, Lawler S. microRNAs in cancer: Biomarkers, functions and therapy[J]. Trends Mol Med, 2014, 20(8):460-469. doi: 10.1016/j.molmed.2014.06.005 pmid: 25027972
[3]	Mitchell PS, Parkin RK, Kroh EM, et al. Circulating microRNAs as stable blood-based markers for cancer detection[J]. Proc Natl Acad Sci USA, 2008, 105(30):10513-10518. doi: 10.1073/pnas.0804549105 pmid: 18663219
[4]	Lena AM, Mancini M, diValCervo PR, et al. microRNA-191 triggers keratinocytes senescence by SATB1 and CDK6 downregulation[J]. Biochem Biophys Res Commun, 2012, 423(3):509-514. doi: 10.1016/j.bbrc.2012.05.153
[5]	Zhang LB, Flygare J, Wong P, et al. miR-191 regulates mouse erythroblast enucleation by down-regulating Riok3 and Mxi1[J]. Genes Dev, 2011, 25(2):119-124. doi: 10.1101/gad.1998711
[6]	Palmieri A, Pezzetti F, Brunelli G, et al. Anorganic bovine bone (Bio-Oss) regulates miRNA of osteoblast-like cells[J]. Int J Periodontics Restorative Dent, 2010, 30(1):83-87.
[7]	Emami N, Mohamadnia A, Mirzaei M, et al. miR-155, miR-191, and miR-494 as diagnostic biomarkers for oral squamous cell carcinoma and the effects of Avastin on these biomarkers[J]. J Korean Assoc Oral Maxillofac Surg, 2020, 46(5):341-347. doi: 10.5125/jkaoms.2020.46.5.341 pmid: 33122459
[8]	Gao Y, Luo TP, Ouyang XW, et al. IGF2BP3 and miR191-5p synergistically increase HCC cell invasiveness by altering ZO-1 expression[J]. Oncol Lett, 2020, 20(2):1423-1431. doi: 10.3892/ol.2020.11693 pmid: 32724385
[9]	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):949-953.
[10]	Saliminejad K, Khorram Khorshid HR, Soleymani Fard S, et al. An overview of microRNAs: Biology, functions, therapeutics, and analysis methods[J]. J Cell Physiol, 2019, 234(5):5451-5465. doi: 10.1002/jcp.27486 pmid: 30471116
[11]	Liu X, Ma XR, Li HY, et al. LINC00472 suppresses oral squamous cell carcinoma growth by targeting miR-455-3p/ELF3 axis[J]. Bioengineered, 2022, 13(1):1162-1173. doi: 10.1080/21655979.2021.2018092 pmid: 35258410
[12]	Ou DM, Wu Y, Zhang JB, et al. miR-340-5p affects oral squamous cell carcinoma (OSCC) cells proliferation and invasion by targeting endoplasmic reticulum stress proteins[J]. Eur J Pharmacol, 2022, 920: 174820. doi: 10.1016/j.ejphar.2022.174820
[13]	张肖. 人生长激素和miR-191/425簇促进乳腺癌的发生发展的机制研究[D]. 合肥: 中国科学技术大学, 2019.
[14]	Kudelova E, Holubekova V, Grendar M, et al. Circulating miRNA expression over the course of colorectal cancer treatment[J]. Oncol Lett, 2022, 23(1):18. doi: 10.3892/ol.2021.13136 pmid: 34868358
[15]	Chen PJ, Pan X, Zhao LW, et al. microRNA-191-5p exerts a tumor suppressive role in renal cell carcinoma[J]. Exp Ther Med, 2018, 15(2):1686-1693. doi: 10.3892/etm.2017.5581 pmid: 29434754
[16]	Chen T, You YN, Jiang H, et al. Epithelial-mesenchymal transition (EMT):A biological process in the development, stem cell differentiation, and tumorigenesis[J]. J Cell Physiol, 2017, 232(12):3261-3272. doi: 10.1002/jcp.v232.12
[17]	Xu WC, Ji J, Xu Y, et al. microRNA-191, by promoting the EMT and increasing CSC-like properties, is involved in neoplastic and metastatic properties of transformed human bronchial epithelial cells[J]. Mol Carcinog, 2015, 54(Suppl 1):E148-E161.
[18]	Kim YE, Won M, Lee SG, et al. RBM47-regulated alternative splicing of TJP1 promotes actin stress fiber assembly during epithelial-to-mesenchymaltransition[J]. Oncogene, 2019, 38(38):6521-6536. doi: 10.1038/s41388-019-0892-5
[19]	Hsu YL, Hung JY, Chang WA, et al. Hypoxic lung cancer-secreted exosomal miR-23a increased angiogenesis and vascular permeability by targeting prolyl hydroxylase and tight junction protein ZO-1[J]. Oncogene, 2017, 36(34):4929-4942. doi: 10.1038/onc.2017.105 pmid: 28436951
[20]	Xia TF, Pan ZG, Zhang J. CircSMC3 regulates gastric cancer tumorigenesis by targeting miR-4720-3p/TJP1 axis[J]. Cancer Med, 2020, 9(12):4299-4309. doi: 10.1002/cam4.v9.12
[21]	Park SY, Jang H, Kim SY, et al. Expression of E-cadherin in epithelial cancer cells increases cell motility and directionality through the localization of ZO-1 during collective cell migration[J]. Bioengineering, 2021, 8(5):65. doi: 10.3390/bioengineering8050065

基因名称	上游引物(5'—3')	下游引物(5'—3')
U6	GGAACGATACAGAGAAGATTAGC	TGGAACGCTTCACGAATTTGCG
miR-191-5P	CCAACGGAATCCCAAAAGCAGC	TGGAACGCTTCACGAATTTGCG
GAPDH	TACTAGCGGTTTTACGGGCG	TCGAACAGGAGGAGCAGAGAGCGA
TJP1	CTGGTGAAATCCCGGAAAAATGA	TTGCTGCCAAACTATCTTGTGA

In vitro study on miR-191-5p targeting TJP1 to regulate the proliferation, invasion and migration of Cal-27 cells

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 11

References 21

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	SHEN Xiaojing, LIU Hairong, LI Hua, LI Linlin, WANG Lewei, YUAN Rongtao, GUO Qingyuan, ZHAO Peng. Effects of total flavonoids of rhizomadrynariae on proliferation and apoptosis of alveolar bone osteoblasts during osseointegration of oral implants [J]. Stomatology, 2023, 43(8): 679-685.
[2]	HU Haiyan, GAO Teng, ZHU Zai'ou, DING Xu, WU Yunong, SONG Xiaomeng. Study on the role of CMTM4 in oral squamous cell carcinoma and its mechanism [J]. Stomatology, 2023, 43(6): 481-487.
[3]	XU Yiping, HUANG Jialing, LIU Zhongbin, YANG Kun, GE Song. Research progress of the relationship between Fusobacterium nucleatum and oral squamous cell carcinoma [J]. Stomatology, 2023, 43(6): 546-551.
[4]	YANG Yuemei,SONG Xiaomeng,WU Yunong. Promotion of cell proliferation by Notch1^P1641S mutation through PI3K/Akt activation in OSCC [J]. Stomatology, 2023, 43(5): 393-399.
[5]	XIAO Tao, HE Yijia, ZHU Yaoping, HAO Fengyao, CHEN Yan, WANG Zhiyong. Study on the mechanism of VEGF inducing tolerogenic dendritic cells in oral squamous cell carcinoma [J]. Stomatology, 2023, 43(3): 204-211.
[6]	JI Huan, LI Meng, YAO Enhui, ZHONG Yi, ZHANG Yuyao, WU Heming, LI Bin. Study on the expression of LncRNA LINC00958 in head and neck squamous cell carcinoma [J]. Stomatology, 2023, 43(11): 968-974.
[7]	XIAO Jinzhi, YU Wei, ZHANG Hao. Berberine hydrochloride inhibits the proliferation of oral squamous cell carcinoma by regulating autophagy [J]. Stomatology, 2023, 43(10): 889-893.
[8]	GAO Zhen,HOU Ruikai,SONG Suocheng,RUAN Jianping. Effect of calcium on biological properties of the ameloblast ALC [J]. Stomatology, 2023, 43(1): 39-45.
[9]	MA Ping, CHENG Luyao, JIN Wulong, WANG Ning, ZHAI Rongping. Expression level and biological function of NCAPD2 in oral squamous cell carcinoma [J]. Stomatology, 2022, 42(8): 688-693.
[10]	WANG Ting, DU Lingqian. Effects of Ghrelin on proliferation and apoptosis of periodontal ligament stem cells [J]. Stomatology, 2022, 42(8): 681-687.
[11]	WANG Jinxin, YI Jie, DING Yujie, ZHONG Yi, SUN Zhida. Expression and role of Claudin-7 in oral squamous cell carcinoma [J]. Stomatology, 2022, 42(7): 604-608.
[12]	HOU Yiming, LI Na, YU Wenqian, CHEN Lei. Research progress of histone modification in oral squamous cell carcinoma [J]. Stomatology, 2022, 42(7): 650-654.
[13]	GUO Zhichen, JING Sili, HU Xiaoyi, ZHANG Zhou, CUI Hao, NA Sijia. Effect of smoking and drinking exposure history on prognosis of patients with oral squamous cell carcinoma at different ages [J]. Stomatology, 2022, 42(6): 521-524.
[14]	JIANG Feng, HE Yao, ZHU Zaiou, DING Xu, YE Jinhai, WU Yunong, SONG Xiaomeng. Effects of knocking down TULP3 on biological behaviors of head and neck squamous cell carcinoma cells [J]. Stomatology, 2022, 42(6): 494-500.
[15]	AINIWAER Ailimaierdan, HUOJIA Muhetaer, WANG Ling. The effect and primary mechanism of transforming growth factor-β3 on the proliferation and osteogenic capability of osteoblasts [J]. Stomatology, 2022, 42(6): 501-507.