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Archive > Volume 29 Issue 3 > 2022,29(3):195-201. DOI:10.3872/j.issn.1007-385X.2022.03.005 Prev Next
Basic Research
Interfering with B7-H4 expression can inhibit proliferation of breast cancer cells by down-regulating E2F family related transcription factors
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Abstract:
Objective: To investigate the effects of interfering with B7-H4 expression on proliferation, apoptosis, cell cycle and expression of downstream molecules in breast cancer cells. Methods: Breast cancer T47D and MCF-7 cells at logarithmic phase were transfected with siRNA specifically targeting B7-H4 (siB7-H4) or its negative control (siNC) by using Lipofectamine TM 2000, namely T47D siNC, T47D-siB7-H4, MCF-7-siNC, and MCF-7-siB7-H4 group, respectively. The efficacy of siRNA interference and its effect on the expression of cyclin D1 were verified by quantitative PCR (qPCR) and Western blotting (WB). The effects of interfering with B7-H4 on cell proliferation, cell cycle and apoptosis of breast cancer cell lines T47D and MCF-7 were detected by CCK-8 assays and flow cytometry, respectively. The effects of interfering with B7-H4 on the expression of E2F family related transcription factors were examined by qPCR. Results: The T47D and MCF-7 cell lines with B7-H4 knockdown were successfully constructed. Compared with the cells in T47D-siNC and MCF-7-siNC groups, the mRNA and protein levels of B7-H4 were significantly decreased, and the proliferation was significantly inhibited in the cells of T47D-siB7-H4 or MCF-7-siB7-H4 groups, accompanied with G1/S cell cycle arrest as well as downregulation of cyclin D1 (all P<0.01); however, there were no statistically significant differences in apoptotic rates (all P>0.05). Compared with the cells in T47D-siNC group, the mRNA levels of E2F1, E2F2, E2F7 and E2F8 in T47D cells decreased in varying degrees after interfering with B7-H4 (all P<0.01); compared with cells in MCF-7-siNC group, the mRNA levels of E2F1, E2F2, E2F3, E2f7 and E2F8 in MCF-7 cells also decreased in varying degrees after interfering with B7-H4 (P<0.05 or P<0.01). Conclusion: Interfering with B7-H4 in breast cancer cells can down-regulate the expression of cyclin D1 and E2F family related transcription factors, leading to cell cycle arrest and inhibition of cell proliferation.
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Clc Number:
R739.7; R730.2
