Inhibition Of Qingre Zaoshi Jiedu Huoxue Formula And Its Seperated Prescriptions On PLCγ1 Signaling Transduction Of RKO Cell Line In Human Colon Cancer

Objective:To screen out effective Chinese medicine which can inhibit the proliferation of human colon cancer cell from clinically commonly used traditional Chinese medicine of treating colon cancer, and compose Qingre Zaoshi Jiedu Huoxue formula, observe the inhibitory effect of Qingre Zaoshi Jiedu Huoxue formula and its three separated prescriptions on colon cancer cell, explore the mechanism and traditional Chinese medicine pathogenesis of Qingre Zaoshi Jiedu Huoxue formula and its three separated prescriptions on colon cancer cell, and provide effective prescription of traditional Chinese medicine treatment of colon cancer. Methods:Literatures of TCM treating colon cancer in China National Knowledge Infrastructure(CNKI) from June 1979 to March 2014 was collected and analyzed, frequency analysis was used to summarize 250 kinds of commonly used Chinese medicine as the preliminary screening herbs. The 250 kinds of herbs were soaked in 95% ethanol and extracted, using the last medicine concentration of 100 ug/ml dealing with colon cancer cell RKO, then observing the morphological change of RKO cell under inverted microscope, after 48 hours, using MTT method to detect and screen out herbs with obviously inhibitory effect to RKO cell, using the orthogonal experimental design to select the curative effect and composing the best medical formula, flow cytometry was used to observe the effect of Qingre zaoshi jiedu huoxue formula and its three separated prescriptions on RKO cell, then western blot method was used to observe the effect of Qingre zaoshi jiedu huoxue formula and its separated prescriptions on PLCγ1 signaling transduction of RKO Cell line in Human Colon Cancer, explore the relationship of TCM pathogenesis of colon cancer and RKO growth signal transduction, looking for effective agent for the treatment of colon cancer. Results: 1 Qingre zaoshi jiedu huoxue formula and its separated prescriptions had obviously inhibitory effect on the proliferation of RKO cell. Compared with the control group, all treatment groups had great effect on cell morphology. The cell morphology of Qingre Jiedu group obviously became small, the number reduced with sparse arrangement; cell morphology in the group of Qingre Zaoshi group became small and the number reduced; cell morphology in the Xingqi Huoxue group became fusiformis and the number reducer to some extent; cell morphology in the whole formula group became small, intercellular substance magnified and cell number reduced greatly. 2 Qingre zaoshi jiedu huoxue formula and its separated prescriptions had obviously inhibitory effect on the proliferation of RKO cell, and the whole formula group had the most significantly effect, and presented dosage-effects and time-effects dependent relationship for the inhibition of RKO cell proliferation with the increasing of medicine concentration and time. 3 Qingre zaoshi jiedu huoxue formula and its separated prescriptions had obviously effect on the cell cycle of RKO cell, cellular percent of G1 stage increased, S stage no obviously change, G2 stage reduced. The results indicated that Qingre zaoshi jiedu huoxue formula and its separated prescriptions inhibited cell proliferation in the G2 stage. 4 EGFR,PLCγ1,PKCα were overexpressed inhuman colon cancer cell RKO. 5 When the cell were treated with IC50 drugs dosage, the protein expression level of EGFR,PLCγ1,PKCαin the cells decreased. Conclusion: 1 Qingre zaoshi jiedu huoxue formula and its separated prescriptions had obviously inhibitory effect on the proliferation of RKO cell, and the whole formula group had the most significantly effect, Qingre Jiedu, Qingre zaoshi and Xingqi huoxue have synergistic effects. 2 The inhibitory effects of Qingre zaoshi jiedu huoxue formula on human colon cancer cell RKO have dose and time dependent relationship. 3 Qingre zaoshi jiedu huoxue formula and its separated prescriptions inhibited cell proliferation in the G2 stage. 4 The results of literature analysis and experimental research show that the colon cancer is closely with the pathogenesis of damp-hot, fire-toxin, qi stagnation and blood stasis.