Fig. 2: PCAT1 promotes ESCC cell proliferation, and knockdown of PCAT1 leads to G2/M phase arrest.

a RT-qPCR analysis of PCAT1 expression in KYSE30 cells knocking down PCAT1. GAPDH was used as an internal reference. sh-NC: shRNA negative control, sh-1/2: shRNA targeting PCAT1. Statistical significance was assessed using one-way ANOVA. *P < 0.05 versus sh-NC. b The growth of KYSE30 cells with PCAT1 knockdown was monitored by the RTCA-MP system. c Colony formation assay was performed using KYSE30 cells with PCAT1 knockdown. Representative images (left) and quantification analysis (right) are shown (n = 3). Statistical significance was assessed using one-way ANOVA. ***P < 0.001 versus sh-NC. d A soft-agar colony formation assay was conducted using KYSE30 cells with PCAT1 knockdown. Representative images (left) and quantification analysis (right) are shown (n = 3). Statistical significance was assessed using one-way ANOVA. ***P < 0.001 versus sh-NC. e RT-qPCR analysis of PCAT1 expression in KYSE150 and KYSE450 cells overexpressing PCAT1. Statistical significance was assessed using two-tailed Student’s t test. **P < 0.01. f, g The growth of KYSE150 and KYSE450 cells with PCAT1 overexpression was monitored by the RTCA-MP system. h, i Colony formation assay was performed using KYSE150 and KYSE450 cells overexpressing PCAT1. Representative images (left) and quantification analysis (right) are shown (n = 3). Statistical significance was assessed using two-tailed Student’s t test. *P < 0.05; **P < 0.01. j The cell cycle distribution of KYSE30 cells with PCAT1 knockdown was determined by PI staining and flow cytometry. One-way ANOVA was used to compare the G2/M phase distribution (n = 3). *P < 0.05 versus sh-NC. k Western blot analysis of cyclin B1, CDC2 and CDK4 protein levels in KYSE30 cells with PCAT1 knockdown. l The paclitaxel sensitivity of KYSE30 cells with PCAT1 knockdown was determined with MTT. Statistical significance was assessed using two-way ANOVA. ***P < 0.001 versus sh-NC