Effect Of WISP2 Gene Overexpression On The Proliferation,invasion And Migration Of Esophageal Cancer Cells Abstract

Objective: To explore the effect and the potential mechanism of WISP2 gene overexpression on proliferation, invasion and migration of esophageal cancer cells.Methods: The plasmid containing WISP2 gene and control were transient transfection into esophageal cancer cell line E109 by Lpofectamin2000, respectively.WISP2 overexpression was verified by quantitative real-time PCR(gene level) and Western blot(protein level) in the group E109-WISP2, E109-blank and E109-empty-vector group. Cell proliferation ability was detected by CCK8 assay and cell invasion and migration were affirmed by matrigel invasion assay and transwell migration assay. The expression of ERK-1/2, Slug and E-cadherin were affirmed in gene level and protein level by q-PCR and Western blot respective.Results: Esophageal cancer cells expressing high levels of WISP2 were successfully obtained. WISP2 expression on gene level and protein level were significantly up-regulated in E109-WISP2 group cells compared with E109-blank and E109-empty-vector groups(p<0.05 for both). Proliferation assay showed that the ability of the E109-WISP2 group cells(0.225±0.003)were reduced compared with E109-blank(0.260±0.001)and E109-empty-vector groups(0.260±0.003)(p<0.05 for both). Invasion assay displayed that the numbers of cells passed through matrigel membrance were significantly reduced in E109-WISP2 group(26.60±4.81) compared with E109-blank(56.00±4.90)and E109-empty-vector groups(53.20±4.32)(p<0.05 for both). Migration assay results indicated that the numbers of cells passed through polycarbonate filters were significantly reduced in E109-WISP2 group(55.20±4.52)compared with E109-blank(99.60±5.78)and E109-empty-vector groups(95.50±4.79)(p<0.05 for both). The expression of ERK-1/2 and slug on gene level and protein level in E109-WISP2 group were reduced than the E109-blank and E109-empty-vector groups(all p<0.05). But the expression of E-cadherin was significantly higher in E109-WISP2 group than the E109-blank andE109-empty-vector groups(all p<0.05).Conclusion: 1. WISP2 gene overexpression could inhibit the proliferation of esophageal cancer cells. 2. WISP2 gene overexpression could inhibit the invasion and migration of esophageal cancer cells. 3. WISP2 gene overexpression could reduced the gene and protein expression of ERK1/2,Slug and up-regulated E-cadherin expression. 4. WISP2 gene could inhibit the the proliferation of esophageal cancer cells by reduced the ERK1/2 signal in the ERK1/2-MAPK signal pathway and inhibit the invasion and migration of esophageal cancer cells by down-regulating the slug then up-regulating E-cadherin expression.