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WT1反义mRNA对卵巢癌SKOV3细胞凋亡诱导的影响。

Effect of WT1 antisense mRNA on the induction of apoptosis in ovarian carcinoma SKOV3 cells.

作者信息

Huo X, Ren L, Shang L, Wang X, Wang J

机构信息

Department of Obstetrics and Gynecology, Beijing General Hospital of Beijing Military Command, Beijing China.

出版信息

Eur J Gynaecol Oncol. 2011;32(6):651-6.

Abstract

PURPOSE

To study the effect of WT1 antisense oligodeoxynucleotide (ASODN) transfection on the proliferation and apoptosis of SKOV3 cells.

METHODS

There were four groups in our study: normal control group, WT1 ASODN group, WT1 SODN group and lipofectamine group. Cell apoptosis was observed by flow cytometry. The effect of WT1 ASODN on cell proliferation was assayed by the MTT method. RT-PCR and Western blot were used to detect the expression level of WT1 mRNA and protein.

RESULTS

The growth of the ovarian cancer cell line SKOV3 became significantly slower and its activity was reduced after being transfected by WT1 ASODN, with the inhibition rate of 49.48%. WT1 antisense phosphorothioate oligonucleotides did not only inhibit cell proliferation, arrest cell cycle at G0-G1 checkpoint and induce apoptosis in SKOV3 ovarian carcinoma cells, but also downregulated WT1 mRNA and protein expression, which contributed to the apoptosis (p < 0.05).

CONCLUSION

WT1 antisense phosphorothioate oligonucleotides could both inhibit the proliferation and induce the apoptosis in SKOV3 ovarin carcinoma cell lines. Antisense oligonucleotides of WT1 may potentially help with the gene therapy of ovarian carcinoma.

摘要

目的

研究WT1反义寡脱氧核苷酸(ASODN)转染对SKOV3细胞增殖和凋亡的影响。

方法

本研究分为四组:正常对照组、WT1 ASODN组、WT1 SODN组和脂质体组。采用流式细胞术观察细胞凋亡情况。采用MTT法检测WT1 ASODN对细胞增殖的影响。采用RT-PCR和Western blot检测WT1 mRNA和蛋白的表达水平。

结果

WT1 ASODN转染后,卵巢癌细胞系SKOV3的生长明显减慢,活性降低,抑制率为49.48%。WT1反义硫代磷酸寡核苷酸不仅抑制SKOV3卵巢癌细胞的增殖,使细胞周期阻滞于G0-G1期并诱导其凋亡,还下调WT1 mRNA和蛋白表达,促进细胞凋亡(p<0.05)。

结论

WT1反义硫代磷酸寡核苷酸可抑制SKOV3卵巢癌细胞系的增殖并诱导其凋亡。WT1反义寡核苷酸可能有助于卵巢癌的基因治疗。

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