Determination of p-coumaric acid in Shuang Bailian mixture and its mechanism of inhibiting the growth of esophageal carcinoma cells / 西安交通大学学报(医学版)

ABSTRACT

【Objective】 To establish a method to determine the content of the effective ingredient PCA (p-coumaric acid) in Shuang Bailian mixture and to investigate its anti-cancer mechanism. 【Methods】 High performance liquid chromatography (HPLC) was used to determine the content of PCA in Shuang Bailian mixture. The CCK8 method was used to detect the antitumor activity of PCA on esophageal cancer cells and the semi-inhibitory concentration of PCA on esophageal cancer cells. Moreover, the nude mice were used to investigate the anticancer effect of PCA. Western blotting was used to detect the expressions of apoptosis-related proteins (cleaved caspase 3, cleaved PARP, Bad, Bcl-2, PI3K, AKT, p-PI3K and p-AKT) in esophageal cancer cells and tumor tissues of nude mice. 【Results】 The concentration of PCA in Shuang Bailian mixture was 16.84 μg/mL. The linear regression equation of PCA was y=204 402x +360 904 (15-40 μg/mL), the RSD of the precision experiment was 2.66%, the RSD of the stability experiment was 2.35%, 3.22%, 1.58%, and 4.08%, respectively. The RSD of the repeatability experiment was 4.01%. The mean value of the recovery rate was 97.83% and the RSD value was 6.16%. CCK8 results showed that the half maximal inhibitory concentration (IC50) of PCA in KYSE30 and KYSE140 cells was 36 μg/mL and 55 μg/mL, respectively. The results of nude mice tumor experiments showed that after 30 days of administration, the tumor xenograft in control mice continued to increase in size, while the cisplatin group, PCA group, and PCA combined with cisplatin administration could effectively inhibit the growth of transplanted tumors in tumor-bearing mice. In addition, Western blotting results showed that compared with the control group, the cisplatin group, PCA group, and PCA combined with cisplatin group could effectively increase the protein expressions of cleaved caspase 3, cleaved PARP, and Bad, but decrease the protein expressions of Bcl-2, p-PI3K, and p-AKT in tumor tissues and KYSE30 and KYSE140 esophageal cancer cells. 【Conclusion】 The concentration of PCA in Shuang Bailian mixture was 16.84 μg/mL, and the HPLC content determination method conditions were sensitive and stable. PCA may have an active anti-tumor effect by regulating the PI3K/AKT/Bcl-2 signaling pathway and inducing apoptosis in esophageal cancer cells.