Apoptotic Influence And Mechanism Research Of Injection Of Prunella Vulgaris On Pancreatic Cancer Cells PANC-1

Objective：The purpose of this study was to investigate the apoptotic influence of IPV (Injection ofPrunella Vulgaris) on human pancreatic cancer cells PANC 1, and to explore the mechanism ofits apoptosis.Methods：1. PANC 1 cells were cultivated in vitro and intervened in by IPV. The experiment wasdivided into six groups, a blank in the control group and five groups with IPV in them.Intervened group were given, 30, 60, 90, 120, 150 mg/ml, five concentration gradients ofIPV. The growth of PANC 1 cells was observed by inverted phase contrast microscopewhen the treatment was completed after 12, 24, 36 and 48 hours, respectively.2. The effect of PANC 1 cells proliferation was studied by MTT. Then drew a survival ratecurve of the different time that PANC 1 cells were treated by IPV, and calculated IC50.3. Used the flow cytometry, the extent of apoptosis of PANC 1 cells was quantified withAnnexin V EGFP and PI under the treatment with different concentrations of IPV after 24hours of intervention.4. The expression of bcl 2 and Bax was examined by RT PCR.Results：1. The observation with inverted phase contrast microscope was showed that pancreaticcancer cells’proliferation was obviously inhibited with the IPV intervened dose increasing.2. The result of MTT was that the effect of IPV inhibition on the growth of PANC 1 cellsbecame gradually stronger with the IPV intervened dose increasing and the extension of theintervened time. These had an obviously dose time dependence (P<0.05). And thedifferences had Statistical Significance. IC50=47.75mg/ml.3. As shown by the flow cytometry, the apoptotic rate of PANC 1 cells could be induced byIPV in the dose dependent manner. And the most of apoptotic changes occurred in earlierperiod of the apoptosis.4. The result of RT PCR indicated that IPV could make PANC 1 cells up regulate theexpression of Bax gene and down regulate the expression of bcl 2 gene after 24hours of theIPV intervention. With the concentration of IPV increasing, the expression of bcl 2 genewas gradually reducing and was down regulated in a dose dependent manner (P<0.05). Theexpression of Bax gene was up regulated in the same manner (P<0.05). The differences of Bax/bcl 2 were both the same as Bax and bcl 2(P<0.05).Conclusion：1. The proliferation of human pancreatic cancer cells can be inhibited by IPV in adose dependent manner.2. IPV can promote the pancreatic cancer cells’apoptosis, and revealing the relationship ofdose dependence.3. This mechanism is that IPV regulates pancreatic cancer cells’gene expression of Bax andbcl 2. The expression of bcl 2 gene was down regulated, and Bax gene was up regulated.Consequently, IPV promotes pancreatic cancer cells apoptosis and inhibition ofproliferation.