Effects of UPF1 on the Proliferation, Migration and Invasion of Breast Cancer Strains MDA-MB-231 and MCF-7

doi:10.13523/j.cb.2107052

China Biotechnology

2022, Vol. 42

Issue (1/2): 58-71 DOI: 10.13523/j.cb.2107052

Orginal Article

Effects of UPF1 on the Proliferation, Migration and Invasion of Breast Cancer Strains MDA-MB-231 and MCF-7

HU Kai,HU Jing,SUN Zi-jiu,LIU Shi-yan,LIAO De-yu,YU Huo-mei,ZHANG Yan(

)

Key Laboratory of Clinical Laboratory Diagnosis, Ministry of Education, College of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China

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Abstract

Objective: To investigate the effects of UPF1 on the proliferation, migration and invasion of human breast cancer cells MDA-MB-231 and its possible mechanism. Methods: The role of UPF1 in breast cancer and the expression level of UPF1 in pan-cancer and breast cancer was evaluated by bioinformatics analysis. Breast cancer cells MDA-MB-231 and MCF-7 were transfected with siRNA. The experiment was divided into two groups: control group (transfected with siRNA negative control) and treatment group (transfected with siUPF1). The mRNA and protein levels of UPF1, MMP9, EMT-related makers were measured by qRT-PCR and Western blot; the proliferation of MDA-MB-231 and MCF-7 cells was detected by CCK-8 assay; the lateral migration ability was studied by wound healing assay; the longitudinal migration and invasion abilities were evaluated by Transwell migration and invasion assays. Results: Bioinformatics analysis showed that UPF1 was highly expressed in breast cancer and associated with immune cell infiltration processes and positively correlated with tumor suppressor genes. UPF1 was also highly expressed in breast cancer cells. After knockdown of UPF1, the mRNA and protein expression levels of UPF1 in MDA-MB-231 and MCF-7 breast cancer cells were significantly decreased (P<0.05, P<0.05). Furthermore, the proliferation, migration and invasion abilities of MDA-MB-231 and MCF-7 cells were significantly enhanced. The mRNA and protein levels of MMP9 and Vimentin were increased, but E-cadherin decreased. Conclusion: UPF1 is highly expressed but plays a cancer-inhibiting role in breast cancer. UPF1 may inhibit the proliferation, migration and invasion of breast cancer MDA-MB-231 and MCF-7 cells by inhibiting the EMT signaling pathway.

Key words： Breast cancer UPF1 Proliferation Migration Invasion EMT

Received: 23 July 2021 Published: 03 March 2022

ZTFLH:

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Corresponding Authors: Yan ZHANG E-mail: yanzhang@cqmu.edu.cn

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	Kai HU
	Jing HU
	Zi-jiu SUN
	Shi-yan LIU
	De-yu LIAO
	Huo-mei YU
	Yan ZHANG

Cite this article:

HU Kai,HU Jing,SUN Zi-jiu,LIU Shi-yan,LIAO De-yu,YU Huo-mei,ZHANG Yan. Effects of UPF1 on the Proliferation, Migration and Invasion of Breast Cancer Strains MDA-MB-231 and MCF-7. China Biotechnology, 2022, 42(1/2): 58-71.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2107052 OR https://manu60.magtech.com.cn/biotech/Y2022/V42/I1/2/58

Table 1 Quantitative real-time PCR primer sequence

Fig.1 UPF1 expression is upregulated in breast cancer tissues (a) Differential expression of UPF1 in pan-cancer in the TNM plot database (b) Differential expression of UPF1 protein in breast cancer and different subtypes of breast cancer (c) Subcellular localization of UPF1 in breast cancer (a) Significant differences by Mann-Whitney U test are marked in red* and labeled after the name of the tissue below the picture

Fig.2 UPF1 was associated with immune infiltration (a) Promoter methylation levels of UPF1 in breast cancer (b) Genomic changes in UPF1 in breast cancer (c) Correlation of UPF1 with NK-cell immune infiltration (d) Correlation of UPF1 with T-cell immune infiltration (e) Correlation of UPF1 with B-cell immune infiltration

Fig.3 UPF1 was positively associated with NMD-related molecules and tumor suppressor gene (a) Correlation of UPF1 with SMG1 (b) Correlation of UPF1 with UPF2 (c) Correlation of UPF1 with ETF1 (d) Correlation of UPF1 with GSPT1 (e) Correlation of UPF1 with APC (f) Correlation of UPF1 with TP53 (g) Correlation of UPF1 with BRCA1 (h) Correlation of UPF1 with BRCA2

Fig.4 UPF1 decreased in breast cancer MDA-MB-231 and MCF-7 cells (a) The expression of UPF1 in MCF-10A,MCF-7,SK-BR-3 and MDA-MB-231 cells were detected by qRT-PCR (b) The expression levels of UPF1 in MDA-MB-231 cells transfected with siRNA-UPF1 were detected by qRT-PCR (c),(d) The expression levels of UPF1 in MDA-MB-231 or MCF-7 cells transfected with siRNA-UPF1 were detected by Western blot * P< 0. 05 (NC vs siUPF1); *** P<0. 001 (MCF-10A vs MCF-7;MCF-10A vs SK-BR-3;NC vs siUPF1); **** P< 0. 000 1 (MCF-10A vs MDA-MB-231)

Fig.5 UPF1 inhibited the proliferation of breast cancer cells (a),(b) The effects of UPF1 on proliferation after silencing UPF1 in MDA-MB-231 cells or MCF-7 cells were determined by CCK-8 assay 24 h: **** P< 0. 000 1 (NC vs siUPF1);48 h: **** P< 0. 000 1 (NC vs siUPF1);*** P< 0. 001 (NC vs siUPF1);72 h: **** P< 0. 000 1 (NC vs siUPF1);*** P< 0. 001 (NC vs siUPF1)

Fig.6 UPF1 inhibited the migration of breast cancer cells (a),(b),(c),(d) The effects of UPF1 on migration after silencing UPF1 in MDA-MB-231 or MCF-7 cells were determined by wound healing test and Transwell migration assay * P< 0. 05 (NC vs siUPF1); ** P<0. 01 (NC vs siUPF1); *** P< 0. 001 (NC vs siUPF1)

Fig.7 UPF1 inhibited the invasion of breast cancer cells (a),(b) The effects of UPF1 on invasion after silencing UPF1 in MDA-MB-231 or MCF-7 cells were determined by Transwell invasion assay ** P<0. 01 (NC vs siUPF1)

Fig.8 UPF1 inhibited the EMT signaling pathway (a),(c) The expression of MMP9 and EMT mRNA in MDA-MB-231 or MCF-7 cells were detected by qRT-PCR (b),(d) The expression of MMP9 and EMT protein in MDA-MB-231 or MCF-7 cells were detected by Western blot qRT-PCR:* P<0. 01 (NC vs siUPF1);** P<0. 01 (NC vs siUPF1);*** P< 0. 01 (NC vs siUPF1) Western blot:* P< 0. 01 (NC vs siUPF1)

Fig.9 Potential pathways of UPF1 regulation of EMT (a) Protein interaction network based on UPF1 and EMT (b) Schematic representation of the UPF1-EMT gene regulation mechanism


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