Biological effects of down-regulating of TIZ gene on ovarian cancer cells in vitro

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Author:
ZHAO Bing-bing(Department of Gynecologic Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China)
ZHANG Wei(Department of Gynecologic Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China)
YANG Zhi-jun(Department of Gynecologic Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China)
WANG Qi(Department of Gynecologic Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China)
LI Li(Department of Gynecologic Oncology, Affiliated Tumor Hospital of Guangxi Medical University, Nanning 530021, China)
Journal Title:
Chinese Journal of Obstetrics and Gynecology
Issue:
Volume 47, Issue 10, 2012
DOI:
10.3760/cma.j.issn.0529-567x.2012.10.013
Key Word:
Ovarian neoplasms; Cell line, tumor; Carrier proteins; Transcription factors;RNA, small interfering; In vitro; TIZ gene

Abstract: Objective To construct TIZ gene RNA interference plasmid,as well as study the biological effects of down-regulating of TIZ gene on ovarian cancer cells.Methods According to the mRNA sequence of TIZ on GenBank,3 pairs of small interference RNA (siRNA) for TIZ gene expression were designed and introduced into SKOV3 cells by liposome transfection reagent; the real-time quantitative PCR (QRT-PCR) was used to detect the interference efficiency and selected the most efficiency siRNA segment;pGPU6/GFP/Neo carrier was used to construct pGPU6/GFP/Neo-siRNA-TIZ-573 restructured interference plasmid,introduced into SKOV3 ccll (sh-TIZ-573/SKOV3).The cell growth curves were made by methyl thiazolyl tetrazolium (MTT) method.The clonogenicity efficiency was observed by clony formation test.The cell cycles were measured by flow cytometry (FCM).The ability of invasion,metastasis and adhesion of ovarian cancer cells were detected by the matrigel invasion assay,transwell migration assay and adhesion assay,respectively.Results QRT-PCR results showed that ovarian cancer SKOV3 cells which transfected by recombinant plasmids pGPU6/GFP/Neo-siRNA-TIZ-573 could down-express TIZ gene.The cell growth curves,FCM and clony formation tests showed that the growth and proliferation of sh-TIZ-573/SKOV3 cells were significantly speeded up compared with the control cells (P < 0.01).There was no significant difference in cell invasion,migration and adhesion between sh-TIZ-573/SKOV3 cells [(48.5 ± 1.7) %,(53.6 ±3.4)%,(64.9 ±5.0)%,respectively] and the control cells (P >0.05).Conclusion The growth of ovarian cancer cells could be speeded up by down-expressing of TIZ gene,which TIZ gene may be play a biological role as a tumor suppressor gene.

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