Wang Daoxin, Liu Yaming, Zhao Wanchen, Wang Ru, Tong Xuhui, Jiang Guojun
School of Pharmacy, Bengbu Medical College/Anhui Engineering Technology Research Center of Biochemical Pharmaceuticals, Bengbu 233030, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2019 Jul 30;39(7):797-803. doi: 10.12122/j.issn.1673-4254.2019.07.08.
To investigate the effect of SRC kinase inhibitor PP2 on the invasion and metastasis of lung cancer A549 cells and explore its molecular mechanism.
MTT assay was used to evaluate the inhibitory effect of PP2 on the proliferation of A549 cells. Cell scratch and Transwell assays were performed to assess the invasion and metastatic capacity of A549 cells after treatment with 1, 2, 4, 8, and 16 μmol/L PP2 for 24 h. Western blotting was used to detect the expressions of connexin43 (Cx43) and MMP-2 in the cells after small interfering RNA (siRNA)-mediated silencing or overexpression of Cx43; the changes in the cell invasion and metastasis in response to PP2 treatment after Cx43 silencing or overexpression were investigated.
MTT assay showed that treatment with PP2 at 2, 4, 8, 16, and 32 μmol/L significantly inhibited the proliferation of A549 cells in a concentration-dependent manner. Treatments with PP2 at 1, 2, 4, 8, and 16 μmol/L for 24 h also concentration-dependently lowered the invasion and metastatic abilities of the cells ( < 0.05). At 4 and 8 μmol/L, PP2 significantly increased the expression level of Cx43 protein and decreased the expression level of MMP-2 protein. Overexpression of Cx43 significantly enhanced the inhibitory effect of PP2 on the cell invasion and metastasis, and Cx43 silencing significantly attenuated the inhibitory effect of PP2 ( < 0.05).
PP2 treatment can suppress the invasion and metastasis of A549 cells possibly by modulating the expression of Cx43.
研究SRC激酶抑制剂PP2对肺癌A549细胞侵袭和转移的影响，并探讨其分子机制。
采用MTT法评估PP2对A549细胞增殖的抑制作用。进行细胞划痕实验和Transwell实验，以评估用1、2、4、8和16 μmol/L PP2处理24小时后A549细胞的侵袭和转移能力。使用蛋白质免疫印迹法检测小干扰RNA（siRNA）介导的Cx43沉默或过表达后细胞中连接蛋白43（Cx43）和基质金属蛋白酶-2（MMP-2）的表达；研究Cx43沉默或过表达后PP2处理对细胞侵袭和转移的影响。
MTT法显示，2、4、8、16和32 μmol/L的PP2处理以浓度依赖的方式显著抑制A549细胞的增殖。用1、2、4、8和16 μmol/L的PP2处理24小时也以浓度依赖的方式降低了细胞的侵袭和转移能力（<0.05）。在4和8 μmol/L时，PP2显著增加Cx43蛋白的表达水平并降低MMP-2蛋白的表达水平。Cx43的过表达显著增强了PP2对细胞侵袭和转移的抑制作用，而Cx43沉默则显著减弱了PP2的抑制作用（<0.05）。
PP2处理可能通过调节Cx43的表达来抑制A549细胞的侵袭和转移。
