The Study On The Induce Effect And Mechanism Of Tumor Cell Apoptosis By 20(S)-ginsenoside Rg3

Malignant tumor is one of the major diseases affecting persons' healthand the invasion and metastasis of malignancy will threatening humans' lifeat last. Traditional therapeutic measure and drugs medicaments are not quiteefficient to certain malignancy. So, It is urgent to find an effective ways tokill the tumors. In China, the herbal has been used as the medicine forthousands of years and it is of great value in fighting against tumors.Ginseng is a kind of valuable herbal medicines from which about fortymonome sapoins can be extracted. And evidently, Ginsenoside-Rg3 in theforty monomer saponins has great effect in the treatment of tumor. But mostof the researches are focused on the tumor's proliferation, invasion andmetastasis And the investigation on the mechanism of apoptosis is limited.Further more, previous researches are applied in the field of digestivesystem and respirative system, It is seldom used in the reproductive system.Therefore, we choose the familiar male reproductive system tumor prostatecarcinoma PC-3M and the familiar female reproductive system tumormammary carcinoma MCF-7 as the experimental object and study on itsfunction as well as the mechanism of 20(s)-ginsenoside Rg3 to inducetumor cell to apoptosis.In this experiment, we adopt MTT assay , AO/EB fluorescencedouble-dye technology and flow cytometry to investigate the effect of20(s)-ginsenoside Rg3 during the process of growing , cell apoptosis andcell cycle of prostate carcinoma PC-3M and mammary carcinoma MCF-7.And then immunocellularchemistry , RT-PCR were utilized to investigatethe expression of apoptotic advanced gene caspase-8 and apoptotic controlgene surviving of PC-3M and mcf-7 by 20(s)-ginsenoside Rg3 from proteinlevel and molecular level.The results were as following: MTT assay displayed that ginsenosideRg3 inhibited the growth of PC-3M and MCF-7. With the growing of Rg3'sconcentration, the inhibitory rate of the experimental group has increasedevidently. The IC50 value of PC-3M is 161.19±0.19μg/ml;the IC50 value ofMCF-7 is 106.61±0.71μg/ml. AO/EB fluorescence double-dye technologyby microscope observation displayed that with the increase of Rg3'sconcentration and the prolong of time. In experimental group of PC-3M andMCF-7, decreasing of normal cells are green fluorescent, increasing ofapoptotic cells are yellow and with chrysordine fluorescent,show cellapoptotic rate is increased. Flow cytometry results display that S stage cellof experiment group evidence increased compared control group(P<0.01),G2/M stage cell evidence decreased compared control group(P<0.01),andevery experiment group have prominent difference(P<0.01),depended onthe Rg3's concentration;appeared apoptotic apex before G1 apex inexperiment group.apoptotic cell number evidence increased comparedcontrol group(P<0.01), depended on the Rg3's concentration.All aboutdisplayed that 20(s)-ginsenoside Rg3 could inhibit DNA synthesize ofprostate carcinoma PC-3M and mammary carcinoma MCF-7, blocking in Sstage, and cell apoptosis. Immunocellularchemistry displayed that with theprolong time by Rg3, caspase-8 is negative in PC-3M and MCF-7, there isno palm yellow grain in cell plasm, cell morphology is normal;experimentgroup of Rg3 24 hour feeble positive, about 20% of cell plasm appearpalm yellow grain ,cell have a little crimple;experiment group of Rg3 48hour strong positive, 90% upwards of cell plasm appear palm brown grain ,and cell show pellet. Survivin is feeble positive in PC-3M andMCF-7,about 25% of cell plasm appear palm yellow grain;experimentgroup of Rg3 48 hour positive, half of cell plasm appear crassitude densepalm yellow grain;experiment group of Rg3 48 hour strong positive,almost all cell plasm have palm brown grain. in experimental group ofPC-3M and MCF-7, the positive cells of caspase-8 and surviving areincreased. RT-PCR displayed that in these two cells by Rg3, the expressionof caspase-8 is increased, so do the surviving.To sum up, from the result of the experiment it is evident that20(s)-ginsenoside Rg3 could inhibit growth of prostate carcinoma PC-3Mand mammary carcinoma MCF-7 and induce these two cells to apoptosis.The possible mechanism of apoptosis is to up-regulate the caspase-8, yetthe result of the experiment does not have the reference on thedown-regulate of survivin.