Figure 7.

Overexpression of Cyclin E2 rescues tigecycline‐induced cell proliferation inhibition in human PDAC cell lines. A, Inhibition rates of CCNE2/vector‐overexpressed AsPC‐1 and HPAC cells after treating with 5 and 10 μmol/L tigecycline, respectively, for the indicated time using MTT assay. B, Western blot assay was used to show the expression of CCNE2, CDK2 and CDK4 after tigecycline treatment. Tubulin was used as control. C, D, The cell cycle was analysed by flow cytometry in CCNE2/vector‐overexpressed AsPC‐1 and HPAC cells after treated with 5 and 10 μmol/L tigecycline, respectively, for 72 h. E, F, Image and quantification of CCNE2/vector‐overexpressed AsPC‐1 and HPAC cells positive for BrdU staining after treating with 5 and 10 μmol/L tigecycline, respectively, for 72 h. Scale bar: 100 μm. G, H, Colony formation was examined by soft agar assay (1000 cells/well) in CCNE2/vector‐overexpressed AsPC‐1 and HPAC cells after treating with 5 and 10 μmol/L tigecycline, respectively, for 21‐28 d. Scale bar = 100 μm. Colony numbers were counted. All experiments were repeated at least three times. All data were shown as the mean ± SD. Student's t test was carried out. *P < .05; **P < .01; ***P < .001. P‐value <.05 was considered as statistically significant