Jin'an Powder Induced Lung Cancer Cell Apoptosis And The Impact Of Survivin Pathway

Lung cancer is the first place in attacking incidences of neoplasm in most countries allover the world.The mortality and attacking incidences increase every year from themid 20 century, currently it has bad therapeutic effects of chemotherapy andmagnitude side effects. To prolong the survival time and improve the living quality ofpatients has become the main purpose and a difficult problem. Recently the researchof herbal monomer has suggested that : the inhibition of most herbal monomers areless obvious than chemo-drug. However, many researchers have proved the herbalprescription has good effects in stabilizing focus, deliminating sub-clinical focus,andimproving mid-survival time, better than chemotherapy. Thus, investigating aprescription that has exact chemical components and obvious curative effects couldshow good curative effects of TCM in important disease.Survivin is the strongestinhibitor of apoptosis so far as it found, and the research focus in apoptosis field.Somedomestic scholar has set about the study of the influences of herbs on it, butsystematic research has not been developed. JAF is the mixture of exact monomersfrom an effective prescription, earlier animal experiments have proved it could inhibitthe tumor growth in naked rats and proliferation of cancer cells in vitro.Ourexperiment is going to investigate the influences of JAF on the Survivin relatedapoptosis pathway. The article is divided into three parts.Part one: The inhibition of JAF on lung cancer cell in vitroPurpose: Study the inhibition of JAF on A549 and H446 lung cancer cell and find thesensitive drug dose and time, and also observe the combined effect of JAF andchemo-drug.Method: By MTS method, study the inhibition of different concentration of JAF,consisting monomers, Vp-16, PDD, JAF combined PDD, and JAF combined Vp-16in different times(12 hours.24 hours.36 hours,48 hours), work out the inhibition rate,and find the sensitive dose and time.Result: JAF show different inhibiting effects and dose-reliance and time reliancetrend.24 hour is a sensitive time. IC50 of A549 cancer cell is 250ug/ml for 24 hours.1C50 of H446 cancer cell is 300ug/ml for 24 hours. The cooperation of chemo-drugand JAF has the best inhibition effects.Conclusion: JAF has obvious anticancer effect on A549 and H446 cancer cells. JAFcan improve the sensitivity of cancer cell to chemo-drug.Part two : The induction of apoptosis of JAF on lung cancer cell in vitroPurpose: Study the induction of apoptosis of JAF on A549 and H446 lung cancer cell,confirm and observe apoptosis phenomenon in morphology.Method: Observe the changes of cyto-membrane, cytoplasm, and karyon of A549and H446 after JAF action under optical microscope, detect the apoptosis rate by flowcytometry machine after herb action, observe apoptosis phenomenon in morphologyby laser confocal machine and HE dying.Result: JAF show obvious induction of apoptosis of A549 and H446 lung cancer cellthat can be observed at cell level.Conclusion: JAF can induce of apoptosis of A549 and H446 lung cancer cell whichhas a dose-reliance and time -reliance trend.Part three: The relationship between induction of apoptosis of JAF on lung cancer andP53, Survivin, and caspase-3 expression.Purpose: Study the influences of JAF on P53, Survivin, and caspase-3 expressionduring induction of apoptosis of A549 and H446 lung cancer cell, explore themolecular mechanism of induction of apoptosis of JAF.Method: Extract general mRNA of A549 and H446 after JAF action, applyfluorescence RT-PCR to detect mRNA expression, and apply immuno-bloting andwestern blotting to detect the protein expression.Result: JAF decrease the Survivin expression and increase the expression of P53 andcaspase-3 that also show dose-reliance and time-relianceConclusion: The induction of apoptosis of A549 and H446 lung cancer cell of JAF isrelated to P53, Survivin, and caspase-3 pathway.