PMC

Cell apoptosis is not involved in the inhibition of tumor growth following QYHJ treatment. (A–C) Cell apoptosis was analyzed by flow cytometry following QYHJ treatment. (D) No statistically significant increase was identified in the apoptosis rate following QYHJ treatment in CFPAC-1, CFPAC-1 control RNAi and CFPAC-1 EphB2 RNAi cells. EphB2, erythropoietin-producing hepatoma cell line-B2; QYHJ, Qingyihuaji formula; FITC, fluorescein isothiocyanate.

QYHJ suppresses tumor growth through inhibiting the cell cycle process. (A–C) Cell cycle was analyzed in CFPAC-1,CFPAC-1 control RNAi and CFPAC-1 EphB2 RNAi cells by flow cytometry following QYHJ treatment. (D) QYHJ treatment blocked the cell cycle in the G0/G1 phase and reduced the S phase proportion in CFPAC-1 and CFPAC-1 control RNAi cells. A statistically significant increase was identified in the G0/G1 phase population (P<0.05) and a decrease was identified in the S phase population in CFPAC-1 and CFPAC-1 control RNAi cells, however, no difference was identified in the CFPAC-1 EphB2 RNAi cells following QYHJ treatment (P<0.05). EphB2, erythropoietin-producing hepatoma cell line-B2; QYHJ, Qingyihuaji formula. ^*P<0.05 compared with the NS group.

High expression of EphB2 predicts a superior response to QYHJ treatment through the EphrinB1-EphB2-CDK6 pathway in CFPAC-1 cells. QYHJ resulted in an unclear change of EphB2 (A and B) mRNA and (C and D) protein level, however, a statistically significant increase was identified in the EphrinB1 (A and B) mRNA (P<0.05, P<0.01 and P<0.05) and (C and D) protein (P<0.05, P<0.05 and P<0.05) level following QYHJ treatment in CFPAC-1, CFPAC-1 control RNAi cells and CFPAC-1 EphB2 RNAi cells. QYHJ also resulted in a statistically significant decrease in the CDK6 (A and B) mRNA (P<0.05) and (C and D) protein (P<0.05) level in CFPAC-1 and CFPAC-1 control RNAi cells (P<0.05), however, no change was identified in CFPAC-1 EphB2 RNAi cells. EphB2, erythropoietin-producing hepatoma cell line-B2; QYHJ, Qingyihuaji formula; CDK6, cyclin-dependent kinase 6; EphrinB1, Eph receptor-interacting B1. ^*P<0.05 and ^**P<0.01 compared with the NS group.

High expression of EphB2 indicates advantageous tumor growth suppression of QYHJ treatment in CFPAC-1 cells. (A) No significant difference was identified in tumor volume following QYHJ treatment in CFPAC-1 EphB2 RNAi cells, however, a significant inhibition was obtained in CFPAC-1 (P<0.05) and CFPAC-1 control RNAi (P<0.05) cells following four weeks of intervention. (B) A statistically significant decrease was identified in tumor weight in CFPAC-1 (P<0.05) and CFPAC-1 Contol RNAi (P<0.01) cells following QYHJ treatment, however, no difference was obtained in CFPAC-1 EphB2 RNAi cells. (C) Tumor weight of nude mice transplanted subcutaneously with CFAPC-1 cells with different levels of EphB2 expression. Each group included nine to 10 nude mice. EphB2, erythropoietin-producing hepatoma cell line-B2; QYHJ, Qingyihuaji formula. ^*P<0.05 compared with the NS group. **P<0.01 compared with the NS group. NS, normal saline.