Experimental Study Of Zhong Jie Feng Induced HCT-8Colon Cancer Cell Apoptosis

Objective we study in vitro and in vivo biological effect of Zhong jie feng for HCT-8colon cancer cell apoptosis.provide theoretical basis for research and treatment of colon cancer before surgery and postoperative adjuvant conventional radiotherapy and chemotherapy.Methods and Materials①In vitro:HCT-8cell line were gained by cultivated in vitro. Set up the low dose group, middle dose group, high dose group and blank control group; Each experimental group according to different doses to join Zhong jie feng injection. Application each tumor cell growth inhibition by tetramethyl azo thiazole blue colorimetric method (MTT) detection in different dose Zhong jie feng injection after24h,48h and72h; cell clone formation experiment is detection cloning in culture12days in different concentrations of Zhong jie feng and calculated cell clone formation rate. Annexin V/PI detection apoptosis of tumor cells and Zhong jie feng injection after24h of reaction..②In vivo:4-week-old BALB/c nude mice and collection of logarithmic phase of HCT-8colon cancer cells, transplanting1x106colon cancer cell planting the subcutaneous of nude mice and set up tumor model. the model rats point blank group, the low dose group, middle dose group and high dose group; each group has10. after3d, we began to observe dynamic tumor growth; Continuous observation for21days, and25days we execute rats and rapid stripping tumors, then measurement of tumor size, Apoptosis was detected with TUNEL. Statistical analysis was conducted for the final results.Results①In vitro:HCT-8colon cancer cells in the lower dosage treatment groups had no obvious growth inhibition. Determined by MTT showed in the same effect time, Middle dose group and high dose group of tumor cell inhibition rate comparative differences are statistically significant (P<0.01), the treatment after48h and72h,the high dose group and low dose group the difference was statistically significant (P<0.05); Cell clone formation experiments showed high dose group of tumor cell growth inhibitory effect is relatively obvious, the dosing group compared with blank control group with significant difference (P<0.05). Annexin V/PI shows, HCT-8cell apoptosis rate also showed a trend of rise with Zhong jie feng increased, blank control group compared with dosing groups had significant difference (P<0.05)②In vivo:After4days, the cancer volume of high dose group and low dose group compared with blank control group the difference was statistically significant (P<0.05); After8days,the cancer volume of high dose group compared with the other three groups have significant difference (P<0.01). TUNEL method detection results show that the high dose group compared with the other three groups that the tumor cell apoptosis index had significant difference (P<0.01), tumor cell apoptosis index has no statistical significance when low dose group compared with blank control group (P>0.05).Conclusion Zhong jie feng have the function of the induction apoptosis of HCT-8for colon cancer tumor cells.