野生型P53诱导的蛋白磷酸酶1对卵巢癌细胞侵袭的影响

The effect of silencing Wip 1 on invasion of ovarian cancer cells

目的:探讨野生型p53诱导的蛋白磷酸酶1(Wip1)与卵巢癌细胞SKOV3侵袭力的关系,揭示Wip1对SKOV3细胞的侵袭和迁移等恶性生物学行为的影响。方法:采用划痕实验、Transwell小室法测定Wip1沉默后SKOV3细胞体外侵袭能力。实时荧光定量反转录-聚合酶链反应(RT-PCR)及Western blot方法检测SKOV3细胞中基质金属蛋白酶2(MMP-2)、基质金属蛋白酶9(MMP-9)、组织金属蛋白酶抑制剂1(TIMP-1)、组织金属蛋白酶抑制剂2(TIMP-2)的表达。结果:细胞迁移距离Wip1-siRNA组(30.33±3.67)短于Control组(59.33±7.69),细胞侵袭到Transwell小室滤膜下细胞数Wip1-siRNA组(17.17±1.84)低于Control组(30.17±3.43),RT-PCR和Western blot检测Wip1-siRNA组MMP-2(0.51±0.01)和MMP-9(0.40±0.03)的表达较Control组(1.00±0.00)降低(均P<0.01);Wip1-siRNA组TIMP-1和TIMP-2的表达与Control组未见差异(P>0.05)。结论:沉默Wip1能够抑制人卵巢癌SKOV3细胞侵袭能力,可能是通过抑制MMP-2、MMP-9的表达实现的。

Objective:To investigate the relationship between wild type p53 induced phosphatase 1(Wip1) and the invasion of ovarian cancer SKOV3 cells,and to explore the effect of Wip1 on the invasion and migration of SKOV3 cells.Methods:Scratch test and transwell chamber method were used to determine the invasion of SKOV3 cells in vitro after Wip1 silencing.The expressions of MMP 2,MMP 9,TIMP 1 and TIMP 2mRNA of SKOV3 cells were detected by real time fluorescent quantitative reverse transcription polymerase chain reaction(RT PCR).The expression of matrix metalloproteinases2(MMP 2),MMP 9,tissue inhibitors of metalloproteinases (TIMP 1) and TIMP 2 protein of SKOV3 cells were determined by Western blot.Results:The cell migration distance in Wip1 siRNA group was significant shorter than that in control group (P <0.01).The number of SKOV3 cells migrated into the transwell chamber membrane in Wip1 siRNA group was significant less than that in control group (P <0.01).When compared with control group,significant reduction was seen in the expression of MMP 2 and MMP 9 in Wip1 siRNA group (P <0.01).Conclusion:Silencing Wip1 can inhibit the invasion of human ovarian cancer SKOV3 cells,which possibly do that by inhibiting the expression of MMP 2 and MMP 9.