关键词:食管肿瘤,细胞运动,细胞增殖,UBE2D3基因

Abstract:Objective To establish a stable transfection of UBE2D3 gene in esophageal carcinoma cell line EC109, and investigate the gene overexpression effects of migration, proliferation, and the sensitivity to cisplatin in the cell line. Methods pEGFP-C1 and pEGFP-UBE2D3 plasmids were transfected into the esophageal carcinoma EC109 strain. G418 screening method was used to select stably transfected cell lines which were named as control group (NC) and overexpression group (OE), respectively. The expression efficiency was verified by RT-PCR and Western blotting, the migration was evaluated by scratch test, and the proliferation and sensitivity of cells to cisplatin were respectively determined by CCK-8 assay and half maximal inhibitory concentration (IC₅₀). Results Relative to EC109 cells, the mRNA expression of UBE2D3 of NC group was 1.010±0.590, that of OE group was 2.026±0.898. The protein expression of UBE2D3 of NC group was 0.923±0.237, OE group was 1.520±0.487, respectively. The expression of mRNA and protein level of UBE2D3 in NC was similar to that in EC109 cells(t=1.005,P＞0.05;t=1.492,P＞0.05), while that in OE was much higher than both above (t=6.682,P<0.05;t=9.150,P<0.05). The migration length of NC group was (0.161±0.062)mm at 12 h, while that of OE group was (0.052±0.007)mm(t=4.370,P<0.05). The migration length of NC group was (0.309±0.071)mm at 24 h, compared with OE group was (0.074±0.012)mm (t=3.644, P<0.05). At 12 h and 24 h, relative to NC group, the migration length of OE group was 0.236±0.051 and 0.241±0.037, respectively, which pointed out that the migration of OE was significantly lower than that of NC. For the 1st day, the proliferation of OE was 0.713±0.220 compared with that in NC (t=13.466,P<0.05), and it was 0.848±0.241 (t=27.241,P<0.05) and 0.742±0.233 (t=15.107,P<0.05) at the 2nd day and 3rd day, respectively. The IC₅₀of OE group was 0.356±0.154(t=4.326,P<0.05) relative to that of NC by disposing with cisplatin at the 1st day, it was 0.445±0.098 (t=5.870,P<0.05) and 0.481±0.096 (t=5.009,P<0.05) at the 2nd day and 3rd day, respectively. The IC50 of OE group was less than that of NC group; when deposed with 5 μg/ml cisplatin, both proliferation of OE and NC group was slower than before. At the 1st day, 2nd day and 3rd day, the proliferation of OE group was 0.606±0.283 (t=8.872,P<0.05), 0.759±0.089 (t=16.771,P<0.05) and 0.569±0.574 (t=7.885,P<0.05) relative to NC group, respectively. Moreover, relative to NC group, fraction surviving of OE group was 0.831±0.323 (t=24.226,P<0.05), 0.875±0.102 (t=33.541,P<0.05) and 0.853±0.359 (t=28.187,P<0.05) at those time points above, respectively. The proliferation of OE was slower compared with that in NC, and the inhibition efficiency of cisplatin on the proliferation of OE was more significant than that of NC. Conclusion UBE2D3 gene overexpressed in esophageal carcinoma cell line EC109 strain may result in the cell′s phenotype inhibition of the migration and proliferation and enhance its sensitivity to cisplatin.

Key words:Esophageal neoplasms,Cell movement,Cell proliferation,UBE2D3 gene