Effects of miR-146a targeting PTP1B on proliferation, migration and invasion of colorectal cancer cells

doi:10.3877/cma.j.issn.2095-2015.2025.04.002

Abstract

Abstract:

Objective

To investigate the effects of microRNA-146a (miR-146a) targeted protein tyrosine phosphatase 1B (PTP1B) on the proliferation, migration and invasion of colorectal cancer cells (HCT-116).

Methods

Human colorectal cancer cell line HCT-116 and normal colon epithelial cell line NCM460 were cultured in vitro, HCT-116 cells were randomly divided into control group, miR-146a NC group and miR-146a mimics group. The expression of miR-146a and PTP1B mRNA in HCT-116 cells was detected by real-time quantitative PCR (RT-qPCR). Cell counting kit (CCK-8) and flow cytometry were used to detect cell survival rate and apoptosis. The migration of HCT-116 cells was detected by scratch test. The invasion of HCT-116 cells was detected by Transwell chamber method. The expression changes of PTP1B, matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blotting. Double luciferase reporter assay was used to verify the targeting relationship between miR-146a and PTP1B.

Results

Compared with those in NCM460 cells, the expression level of miR-146a in HCT-116 cells were significantly lower, and the expression of PTP1B mRNA was significantly higher (P<0.05). Dual luciferase assay confirmed that miR-146a has a targeted binding site with PTP1B. Compared with those in control group and miR-146a NC group, the miR-146a expression level and apoptosis rate of HCT-116 cells in miR-146a mimics group were significantly higher (P<0.05), the cell survival rate, scratch migration rate, invasion rate, expression levels of PTP1B mRNA and protein, MMP-2 and MMP-9 protein were significantly lower (P<0.05).

Conclusion

Overexpression of miR-146a may inhibit malignant behavior of colorectal cancer cells by targeting negative regulation of PTP1B expression.

Key words: Colorectal cancer, miR-146a, Protein tyrosine phosphatase 1B, Proliferation, Migration, Invasion

Xujuan Lu, Zhenxing Kong. Effects of miR-146a targeting PTP1B on proliferation, migration and invasion of colorectal cancer cells[J]. Chinese Journal of Digestion and Medical Imageology(Electronic Edition), 2025, 15(04): 294-299.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhxhbyyxzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.2095-2015.2025.04.002

https://zhxhbyyxzz.cma-cmc.com.cn/EN/Y2025/V15/I04/294

Figures/Tables 13

References 21

[1]	Matsuda T, Fujimoto A, Igarashi Y. Colorectal Cancer: Epidemiology, Risk Factors, and Public Health Strategies[J]. Digestion, 2025, 106(2): 91-99.
[2]	Leowattana W, Leowattana P, Leowattana T. Systemic treatment for metastatic colorectal cancer[J]. World J Gastroenterol, 2023, 29(10): 1569-1588.
[3]	Wu Y, Xiao Y, Ding Y, et al. Colorectal cancer cell-derived exosomal miRNA-372-5p induces immune escape from colorectal cancer via PTEN/AKT/NF-κB/PD-L1 pathway[J]. Int Immunopharmacol, 2024, 143(Pt 1): 113261.
[4]	Zhuang J, Shen L, Li M, et al. Cancer-Associated Fibroblast-Derived miR-146a-5p Generates a Niche That Promotes Bladder Cancer Stemness and Chemoresistance[J]. Cancer Res, 2023, 83(10): 1611-1627.
[5]	陈辉国, 周亚夫, 颜建华, 等. 肺癌miR-146a表达及其对细胞增殖,侵袭及迁移的影响[J]. 临床肿瘤学杂志, 2020, 219(3): 37-42.
[6]	Sun X, Cui S, Fu X, et al. MicroRNA-146-5p promotes proliferation, migration and invasion in lung cancer cells by targeting claudin-12 [J]. Cancer Biomark, 2019, 25(1): 89-99.
[7]	Gui H, Nie Y, Yuan H, et al. The combination of a PTP1B inhibitor, TNFR2 blocker, and PD1 antibody suppresses the progression of nonsmall cell lung cancer tumors by enhancing immunocompetence [J]. Oncol Rep, 2024, 52(5): 149.
[8]	余红璐, 任亚君, 熊枝繁. PTPlB和Sam68在结直肠癌中的表达及其意义[J]. 山西医科大学学报, 2017, 48(6): 596-601.
[9]	Yang Q, Zhang L, Zhong Y, et al. miR-206 inhibits cell proliferation, invasion, and migration by down-regulating PTP1B in hepatocellular carcinoma[J]. Biosci Rep, 2019, 39(5): 1-14.
[10]	Abedizadeh R, Majidi F, Khorasani HR, et al. Colorectal cancer: a comprehensive review of carcinogenesis, diagnosis, and novel strategies for classified treatments[J]. Cancer Metastasis Rev, 2024, 43(2): 729-753.
[11]	Moorman A, Benitez EK, Cambulli F, et al. Progressive plasticity during colorectal cancer metastasis[J]. Nature, 2025, 637(8047): 947-954.
[12]	Panoutsopoulou K, Liu Y, Avgeris M, et al. Repression of miR-146a in predicting poor treatment outcome in triple-negative breast cancer[J]. Clin Biochem, 2023, 114: 43-51.
[13]	邢智伟, 高紫玉, 高雅楠, 等. LncRNA MALAT1通过靶向miR-146a调节PI3K/Akt信号通路影响胃癌细胞的增殖、迁移和侵袭[J]. 沈阳药科大学学报, 2024, 41(5): 581-589, 618.
[14]	Noorolyai S, Baghbani E, Aghebati Maleki L, et al. Restoration of miR-193a-5p and miR-146 a-5p Expression Induces G1 Arrest in Colorectal Cancer through Targeting of MDM2/p53[J]. Adv Pharm Bull, 2020, 10(1): 130-134.
[15]	Chen J, Jiang Q, Jiang XQ, et al. miR-146a promoted breast cancer proliferation and invasion by regulating NM23-H1[J]. J Biochem, 2020, 167(1): 41-48.
[16]	梁雪, 吕磊, 钱立庭, 等. miR-146a-5p靶向IRAK1, TRAF6抑制小细胞肺癌耐药[J]. 安徽医科大学学报, 2020, 55(3): 333-339.
[17]	Liang S, Tran E, Du X, et al. A small molecule inhibitor of PTP1B and PTPN2 enhances T cell anti-tumor immunity[J]. Nat Commun, 2023, 14(1): 4524.
[18]	Huang WC, Yen JH, Sung YW, et al. Novel function of THEMIS2 in the enhancement of cancer stemness and chemoresistance by releasing PTP1B from MET[J]. Oncogene, 2022, 41(7): 997-1010.
[19]	Li Y, Luo H, Lin X, et al. Triterpenes of Prunella vulgaris Inhibit Triple-Negative Breast Cancer by Regulating PTP1B/PI3K/AKT/ mTOR and IL-24/CXCL12/CXCR4 Pathways[J]. Int J Mol Sci, 2025, 26(5): 1959.
[20]	Zhu S, Bjorge JD, Fujita DJ. PTP1B contributes to the oncogenic properties of colon cancer cells through Src activation[J]. Cancer Res, 2007, 67(21): 10129-10137.
[21]	Hoekstra E, Das AM, Swets M, et al. Increased PTP1B expression and phosphatase activity in colorectal cancer results in a more invasive phenotype and worse patient outcome[J]. Oncotarget, 2016, 7(16): 21922-21938.

基因	上游引物(5′-3′)	下游引物(5′-3′)
miR-146a	TGTTGCTGAAGAGCTCTGGTAC	TCGTCTGATGATGTCGAATGTAC
PTP1B	GAGTTCG AGCAGATCGACAAGTC	TAGGAAGCTTGGCCACTCTACAT
U6	CTCGCTTCGGCAGCACA	AACGCTTCACGAATTTGCGT
GAPDH	TGGGTGTGAACCATGAGAAGT	TGAGTCCTTCCACGATACCAA

基因	上游引物(5′-3′)	下游引物(5′-3′)
miR-146a	TGTTGCTGAAGAGCTCTGGTAC	TCGTCTGATGATGTCGAATGTAC
PTP1B	GAGTTCG AGCAGATCGACAAGTC	TAGGAAGCTTGGCCACTCTACAT
U6	CTCGCTTCGGCAGCACA	AACGCTTCACGAATTTGCGT
GAPDH	TGGGTGTGAACCATGAGAAGT	TGAGTCCTTCCACGATACCAA

组别	miR-146a	PTP1B mRNA
正常组	1.03±0.19^a	1.01±0.18^a
对照组	0.54±0.09	2.16±0.20
miR-146a NC组	0.53±0.08	2.03±0.19
miR-146a mimics组	0.91±0.11^ab	0.87±0.10^ab
F值	25.068	91.293
P值	<0.001	<0.001

组别	miR-146a	PTP1B mRNA
正常组	1.03±0.19^a	1.01±0.18^a
对照组	0.54±0.09	2.16±0.20
miR-146a NC组	0.53±0.08	2.03±0.19
miR-146a mimics组	0.91±0.11^ab	0.87±0.10^ab
F值	25.068	91.293
P值	<0.001	<0.001

组别	荧光素酶相对光活性
PTP1B-3'UTR-WT+miR-146a NC组	0.85±0.10
PTP1B-3'UTR-WT+miR-146a mimics组	0.24±0.03^a
PTP1B-3'UTR-MUT+miR-146a mimics组	0.76±0.06
PTP1B-3'UTR-MUT+miR-146a NC组	0.78±0.05
F值	111.471
P值	<0.001

Please choose a citation manager

Content to export