Effects Of Exogenous PDCD4 Transfection On The Growth And Apoptosis Of Eca109 Esophageal Cancer Cell Lines

ObjectiveTo study the effect of PDCD4 on the morphology, proliferation and apoptosis of the esophageal cancer cells, we transfected the exogenous PDCD4 into Eca109 esophageal cancer cells compared with the reference group of PLXSN transfected cells and non-transfected cells. Providing a theoretical basis for experimental studies and clinical gene therapy according to this study.Methods1. PLXSN or PDCD4 recombinant plasmid was transfected into the Eca109 esophageal cancer cell lines. And RT-PCR and Western blot methods were employed to detect the transfection expression of PDCD4 in esophageal cancer cell lines.2. We detected the morphological changes of the Eca109 esophageal cancer cells after transfected by PDCD4 through fluorescence inverted microscope.3. We counted the esophageal cancer Eca109 cells by microscope and analyzed the effect of PDCD4 on cells growth and proliferation after 48 hours of the transfection.4. We used flow cytometry to analysis and compare the apoptosis about three groups of Eca109 esophageal cancer cells.Results1. PDCD4 was successfully transfected and stably expressed in the Eca109 esophageal cancer cells. Under fluorescence microscope, green fluorescence was seen in the Eca109 esophageal cancer cells transfected by PLXSN or PDCD4 recombinant plasmid, whereas non-transfected Eca109 esophageal cancer cells cannot be seen fluorescence. Besides, PDCD4mRNA and protein can be potently expressed in the cells of PDCD4 recombinant expression vector by RT-PCR and Western blot method, which was shown that PDCD4 gene was successfully transfected and stably expressed in the Eca109 esophageal cancer cells.2. PDCD4 can change themorphology of the Eca109 esophageal cancer cells. The number and morphology of the cells were significantly different from the normal of the non-transfected cells. The shape of Eca109 esophageal cancer cells transfected by PDCD4 was became round and atrophy, even some cells of which were dead.3. The expression of PDCD4 have obvious influence on the growth of Eca109 esophageal cancer cells. The count of cells transfected by PDCD4 was (5.02±0.29) ×105, which was obviously less than the untreated control (7.67±0.44)×105and the mock-transfected groups (6.81 ±0.33) x 105with significant differences(P<0.05).The untreated control and the mock-transfected groups had no difference (P>0.05)4. The expression of PDCD4 can induce the apoptosis of the Eca109 esophageal cancer cells. Apoptosis rate of the Eca109 esophageal cancer cells transfected by PDCD4 (38.07%±1.08%) was significantly more than the control groups of the negative (19.57%±2.6%) and mock-transfected (26.9%±0.61%) with obvious differences (P<0.05)Conclusions1. PDCD4 gene was successfully transfected and stably expressed in the Eca109 esophageal cancer cells.2. After the PDCD4 gene was transfected into human esophageal cancer Eca109 cells, the cell growth and proliferation was inhibited, suggesting that PDCD4 can inhibit the growth of Eca109 esophageal cancer cells.3. The expression of PDCD4 can promote the apoptosis of Eca109 esophageal cancer cells, which suggest that promoting apoptosis may be a tumor suppressor mechanism of PDCD4.