Experimental Study Of The Apoptosis Inducing Effect Of Histamin-Indomethacin On Lewis Lung Cancer In Mice

ObjectiveBoth histamin and indomethacin(HA-IN) have the anti-tumor function,which are combinded together by the special method.Using the tending-tumor effect of indomethacin and histamine transshipping onto tumor focus,in order to play indomethacin the anti-tumor effect.The experiment studies the effect of inhibition and apoptosis on indomethacin and histamine-indomethacin to Lewis lung cancer in mice with flow cytometry,radio-immunity method ect,and it provides an important theory platform for clinical.Material and MethodsLewis lung cancer were inoculated with tumor in left armpit subcutaneously, when the tumor grew to 1cm in diameter,the tumor bearing mice were divided into 5 groups:3.0 mg/kg indomethacin(IN) group,3.5 mg/kg indomethacin group,3.0 mg/kg histamin-indomethacin(HA-IN) group,3.5 mg/kg histamin-indomethacin (HA-IN) group and saline control group.All groups were administrated the drug once everyday by pouring into stomach observed 10 days continuously.By observed the tumor volume of Lewis daily after treatment,the tumor inhibitory rate was calculated and the tumor inhibition curves was measured,which reflect degree of inhibition of tumor in the different dose groups.By flow cytometry detection with ANNEXINV-FITC+PI double-staining methods and PI single standard labeling,the apoptotic peak and the apoptosis rate of tumor cells for different treatment groups were determined,in order to reflect the induction of apoptosis in tumor to treatment groups. The content of rumor necrosis factor(TNF-α) levels was estimated by the radio-immunity method,and the effect of the different dose groups on TNF-αwas the quantitative analysis.The effect of anti-tumor on different treatment groups was reflected indirectly by the high or low content of TNF-α.Finally,from the comprehensive results evaluated different treatment groups in many respects,especially the final result of histamine-indomethacin anti-tumor.ResultBy observed the tumor volume and the quality of Lewis to different dose groups daily,the tumor inhibitory rate was calculated according to the volume formula, different dose groups have significant difference(P< 0.05) compared with the control group,which inhibition effects were evident at high dosage HA-IN.The tumor was labeled by ANNEXINV-FITC+PI double-staining methods and PI single standard labeling,flow cytometry determined that the apoptosis rate of tumor cells for the different groups(3.0mg/kg HA-IN group,3.5mg/kg HA-IN group,3.0mg/kg IN group and 3.5mg/kg IN group) were respectively 24.7%,34.5%,21.2%,and 23.4%.Flow cytometry with PI single standard labeling determined that the different groups appeared an apoptotic peak before G₀/G₁ phase,and the apoptotic peak of HA-IN groups appeared earlier than that of IN groups,and the number of apoptotic cells was more,especially was more obvious in 3.5 mg/kg HA-IN group.The content of TNF-αlevels measured by the radio-immunity method arrived that the content of TNF-αlevels is significantly increased when compared with before treatment for the different treatment groups of Lewis.In different dosage groups,the content of TNF-αat high dosage groups were significantly higher than those at low dosage groups,so the content of TNF-αlevels showed in a dosage-dependant manner.In addition,compared the different dose HA-IN groups to the different dose IN groups,the content of TNF-αlevels of 3.5mg/kg HA-IN group was significant different(P<0.05) compared with other treatment groups.But the content of TNF-αlevels of 3.0mg/kg HA-IN group was no significant different(P<0.05) compared with different dose IN groups.The results showed that the different dose groups the apoptosis rate,the apoptosis rate and the content of TNF-αlevels were significantly different compared with the control group, and high dose groups HA-IN has obvious effect compared with other treatment groups whether on tumor inhibition and on inducing tumor cell apoptosis and promoting the releasing of TNF-αto induce indirectly antitumor immunity effect.So HA-IN provides a new researching field for carcinoma radiotherapy in clinic. ConclusionsHistamin and indomethacin were combined together by the special method,and were used in the experimental study of antitumor.Various indexes were comprehensively determined to show that HA-IN groups can resist the growth of the tumor and promote cell apoptosis.Among the effect of 3.5mg/kg HA-IN groups was greater,and it was idea than that of single IN.So,we have reasons to think HA-IN is effective in anti-tumor field as a drug and provides an important theoretical platform for the carcinoma radiotherapy in clinic.