| Objective: In the thesis, the different presciptions includesqudengfang,huatanfang and jiedufang which result from differentpreparation methods and their effects on human esophageal cancer cellEC9706proliferation inhibition, cell cycle and protein PLCγ1, PKCα, PI3Kare researched in order to get knowledge of the presciptions’s curemechanism of esophageal cancer, to seek an effective method for preparingdrugs, and explore the molecular mechanismMethods: Alternative medicine was distilled by alcohol and by boilingwith alcohol depositing respectively, MTT assay was used to filter out theappropriate drugs whose inhibition rates is higher and extrate cream ismore.these drugs are divided into qudengfang,huatanfang andjiedufang,according to the " pharmacy ". orthogonal test was designed toselect the best drug combinations; cell EC9706was acted respectively withqufengfang,huatanfang, jiedufang,andqufenghuatanjieduhefang and thenwere observed to the effects of each group on its growth and proliferation ofthe state under an inverted microscope; flow cytometry was used todetect the role and influence of each group drugs on cell’s growth cycle;Wertern blot was used to test the huoxuefang、 jiedufang、 huatanfang andhefangzu’s effects on protein expression of PLCγ1, PKCα, PI3K in esophageal cancer cell EC9706.Results:①of the drug candidates, the drugs whose extrate cream ismore, apparent efficacy and inhibition rate detected by MTT assay forinhibition of human esophageal cancer cell EC9706is higher were selected.changshan， chanpi， distilled by boiling with alcohol depositing chantui，tujingpi， zaojiao， banmao distilled by alcohol were included. According tothe quantity of extrate cream, the order is chanpi， zaojiao， banmao，tujingpi， changshan， chantui. And according to the inhibition rates, theorder is zaojiao， changshan， chanpi， banmao， chantui， tujingpi。②thedrugs were initially divided into qufengfang， huatanfang and jiedufangbased on the results of drug screening and drug efficacy.the optimumcombination of qufenghuatanjieduquanfang was compeled based onorthogonal text design.③human esophageal cancer cell EC9706wasintervened by qufenghuatanjiedufangjiqichaifang, cell morphology changedsignificantly acted with each group drugs was observed under an invertedmicroscope; according to the different groups drugs in different doses andduration of action of the cell line, dose-dependent and time-dependentmanner was present, the order of inhibition rate washuatanjieduhuoxiefang、 huatanfang、 jiedufang、 qufengfang.④all of thecell ratios have declined with each group of therapies in G0/G1phase,especially quanfengzu; the cell ratio of each group drugs has declined in Sphase,and a further decreased in the cell ratio with qufengzu and jieduzu;the cell ratio of each group drugs has increased in G2/M phase, andincreased significantly with qufengzu.⑤PLCγ1, PKCα, PI3K proteins inhuman esophageal cancer cell EC9706show high expression status detectedby Wertern blot, the high expression of proteins was inhibited in differentdegrees with each group drug, in which the most significant effect of the mis quanfang, followed by huatan and jiedufang.Conclusion:①the proliferation of esophageal cancer cell can beinhibited significantly with qufenghuatanjiedujiqichaifang and theinhibitation can be enhanced with qufeng、huatan、jieduyaowu which can bemutual compatibility and synergistic.②esophageal cancer cells werearrested in S phase with huatanzu and quanfangzu, in G2/M phase withqufengzu and jieduzu, and this is an important factor in the inhibition of cell proliferation of esophageal cancer.③PLCγ1, PKCα, PI3K proteinsexpression status in esophagea cell were adjusted and this is key mechanismfor inhibiting the proliferation of esophageal cancer cells withqufenghuatanjiedufangjiqichaofang.④from the effects ofqufenghuatanjiedufangjiqichaifang, it can be speculated that “fengxie，tan， du”can be related to PLCγ1, PKCα, PI3K protein expression statusclosely. |
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