Silencing PAK5 by CRISPR/Cas9 Inhibits Breast Cancer Cell Proliferation and Promotes Cell Senescence
HU Bing-Tao, XING Yao, LI Yang, LI Feng*
（Department of Molecular Cell Biology, College of Life Sciences, China Medical University,Key Laboratory of Medical Cell Biology, Ministry of Education, Shenyang 110122, China）
p21-activated kinase 5 (PAK5), a kind of the PAK family of serine/threonine kinases, modulates various cellular processes, including cytoskeletal remodeling, cell proliferation, migration and metastasis. Although studies have suggested that PAK5 is a key regulator of breast cancer progression, the link of PAK5 to senescence has not been reported yet. In this study, the CRISPR/Cas9 lentivirus infection method was used to construct two PAK5 stably knockdown cell lines of human breast cancer (MDA-MB-231 and BT474). The effect of PAK5 knockdown on the proliferation of breast cancer cells was detected by CCK-8 and clone formation assays. The effect of PAK5 knockdown on cell cycle was detected by flow cytometry. The effect of PAK5 knockdown on cell senescence was observed by β-galactosidase staining. Western blotting was performed to detect the expression of senescence-related proteins including p53, p21 and p16 in breast cancer cells. Cells were treated by CHX and MG132 to explore the possible mechanism of PAK5 regulating p53 protein expression. The results showed that knockdown of PAK5 inhibited cell proliferation and blocked cells in the G0/G1 phase (P＜0.05); Moreover, knockdown of PAK5 promoted cell senescence by upregulating the expression of cellular senescence-related proteins p53, p21 and p16 (P<0.05). Further studies have proved that PAK5 may degrade p53 by ubiquitination. These findings suggest that knockdown of PAK5 can inhibit breast cancer cell proliferation and promote cell senescence, providing potential targets for the treatment of breast cancer.
胡冰涛，邢瑶,李洋， 李丰. CRISPR/Cas9沉默PAK5抑制乳腺癌细胞增殖并促进细胞衰老[J]. 中国生物化学与分子生物学报, 2021, 37(4): 495-503.
HU Bing-Tao, XING Yao, LI Yang, LI Feng. Silencing PAK5 by CRISPR/Cas9 Inhibits Breast Cancer Cell Proliferation and Promotes Cell Senescence. Chinese Journal of Biochemistry and Molecular Biol, 2021, 37(4): 495-503.
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