Effect of ITCH on proliferation, invasion and apoptosis of lung cancer A549 cells

LAN Bing; ZHANG Dongwei; KONG Jinliang; ZHONG Jiajiang; HE Chanjuan

doi:10.3969/j.issn.1009-0460.2019.01.003

Abstract：Objective To investigate the expression of ITCH in lung cancer cells and the effect of ITCH silencing on proliferation, invasion and apoptosis of lung cancer cells A549 and its possible mechanisms. Methods Western blotting was used to detect the expression of ITCH protein in human normal lung epithelial cell lines BEAS-2 B and human lung cancer cell lines A549 and Calu3.Si ITCH (si ITCH group) and si NC (si NC group) were transfected into A549 cells by PEI transfection reagent, respectively. CCK-8 method, Transwell test and flow cytometry were used to detect the changes of cell proliferation, invasion and apoptosis in the two groups. The protein expression of ITCH, Bcl-2, Bax, Cyclin D1, MMP-2, MMP-9, β-catenin and E-cadherin in the two groups were detected by Western blotting assay. Results The expression level of ITCH protein in human lung normal epithelial cell line BEAS-2 B was 0. 98 ± 0. 043, significantly lower than that in Calu3 cell line (1. 92 ± 0. 073) and A549 cell line (2. 74 ± 0. 151), and the difference was statistically significant (P ＜ 0. 05). The cell proliferation rate in si ITCH group was 1. 97 ± 0. 021 at 96 h, which was significantly lower than that in si NC group (3. 72 ± 0. 232), and the difference was statistically significant (P ＜ 0. 05). The apoptotic rate in si ITCH group was (43. 2 ± 1. 52) ％, which was significantly higher than that in si NC group (2. 3 ± 0. 32) ％. The difference was statistically significant (P ＜ 0. 05). The number of perforating cells in si ITCH group was 297. 2 ± 21. 21, which was significantly lower than 601. 2 ± 3. 21 in si NC group (P ＜ 0. 05). Compared with the si NC group, the expression levels of Bcl-2, Cyclin D1, MMP-2, MMP-9 and β-catenin in si ITCH group decreased, while the expression levels of Bax and E-cadherin increased. Conclusion Silencing ITCH inhibits lung cancer proliferation, invasion and induces apoptosis by regulating signaling pathways such as MMP, Cyclin D1 and Bcl-2/Bax.