Center for Research and Development of Antineoplastic Drugs, Tianjin Institute of Medical and Pharmaceutical Science, Tianjin (300020), China;Anorectal Department of Traditional Chinese Medicine Hospital, Binhai New District, Tianjin (Tianjin 300451)
Abstract:
Objective To investigate the effect of Fuzheng Jiedu Quyu formula (Fuzheng formula) combined with Oxaliplatin (L-OHP) on human colon cancer HT-29 cells in nude mice. Methods Mouse model of transplanted colon cancer HT-29 in nude mice was constructed. After tumor formation, they were randomly divided into 4 groups: blank group, Fuzheng formula group, L-OHP group and Fuzheng formula combined with L-OHP (Fuzheng+L-OHP) group, n=10 in each group. In the blank group, mice received normal saline injections throughout the entire 21 d study period, Fuzheng formula groupreceived intragastric administration of Chinese herb (50 g/kg), and L-OHP group were intraperitoneally (ip) given L-OHP (10 mg/kg) once. Fuzheng+L-OHP group were given intraperitoneal injection of L-OHP (ip) once, and intragastric (ig) administration of Fuzheng formula daily for 21 consecutive days. The effect of Fuzheng formula combined with L-OHP on the tumorinhibition rate of subcutaneous transplanted tumor was observed. The expression levels of cyclin D1, CDK4, PCNA and P21 in transplanted tumor were observed by RT-PCR and Western blotting, and the changes of each group were compared. Results The tumorinhibiting rates were 6 . 7 %, 26 . 7 % and 46 . 7 %, respectively in Fuzheng formula group, L-OHP group and Fuzheng+L-OHP group. All of these can inhibit the growth of colon cancer HT-29 transplantation tumor. The spleen index in blank group (0.41±0.04), combined group (0.45±0.11), and Fuzheng formula group (0.47±0.05) were higher than that in L-OHP group (0.40±0.11), and the Fuzheng formula group was significantly higher than that of blank group (P < 0 . 01). Results of RT-PCR indicated that, compared with blank group, the expression levels of cyclin D1, CDK4 and PCNA mRNA were down-regulated in Chinese medicine group, L-OHP group and Fuzheng+L-OHP group (P<0.05, P<0.01), and the expression level of P21 was up-regulated (P<0.01). As compared with the L-OHP group, the expression level of PCNA mRNA down-regulated in Fuzheng+L-OHP group (P<0.01), and the expression level of P21 was up-regulated (P<0.01). Results of Western blotting showed that, compared with blank group, the expression levels of cyclin D1, CDK4 and PCNA protein were down-regulated in Fuzheng formula group, L-OHP group and Fuzheng+L-OHP group (P<0.05, P<0.01), but the expression level of P21 was up-regulated (P<0.05); compared with the L-OHP group, the expression levels of Cyclin D1 and PCNA mRNA were down-regulated in Fuzheng+L-OHP group (P<0.01), and the expression of P 21 was up-regulated (P <0.01). Conclusion Fuzheng formula, L-OHP and their combination can inhibit the growth of transplanted tumors in mice with colon cancer and the combination of the two drugs can not only enhance the effect of chemotherapy, but also inhibit the toxicity of L-OHP to some extent. The mechanism may be related to the expression of cyclin D1, CDK4, PCNA and P21.
