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Rudiments of GSTθ and Cancer Dependability

Author: QiaoHuiPing
Tutor: FanSaiJun
School: Suzhou University
Course: Radiation Medicine
Keywords: GSTθ HeLa cells HCT116 cells cell growth cell cycle invasion radiosensitivity
CLC: R73-3
Type: Master's thesis
Year: 2009
Downloads: 35
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Abstract

Cancer is one of the most common malignancies jeoparding the health all over the world. The incidence of cancer in our country is a million and six hundred thousand per year. Cancer is overtopping heart and cerebrovascular disease becoming the primary disease causing death. Therefor, prevention and cure of cancer become the first topic to follow with interest.In the process of tumorigenesis, abnormity of metabolism lead to malignant biological behaviour directly. The mainspring of that is the key enzyme’s anomal-express which is the consequence of gene regulation and express. Thus tumor related gene and enzymology has significance in the study of tumor. We should search for enzyme mark as clinical diadynamic criteria and drug treatment target.There are many investigations demonstrated that GST is one of the inactivate enzyme of cancerogen, and GST may be the organism mark of precancerosis.Our objective is to study the effect of GSTθincreased expression of the cell growth, cell cycle and radiosensitivity were used in this study.We select two cancer cell lines, HeLa and HCT116. Increased expression of GSTθin the cells was performed by stable transfection with a mammalian pcDNA3-GSTθexpression vector. GSTθmRNA and protein were detected by RT-PCR and Western blot assays, respectively. Cell count assays were applied for evaluation of cell growth; MTT assay was used to detect the dependence of tumor cells to serum levels; the flow cytometry was used to decide the diversity of cell cycle; and colony formation assay was employed to determine cell survival following irradiation. Flow cytometry was used to analyze cell cycle arrest.We also obversed the change of invasion through the scarification experiment.The results were summarized:The expression of GSTθin HeLa and HCT116 cells was undetectable or detected at a low level. Increased expression of GSTθsignificantly caused a G2/M arrest and reduced cell growth, more striking in HCT116 than in HeLa. The growth of cells containing GSTθtransfection became to be more dependent on the serum concentration in HCT116 cells, not in HeLa cells. GSTθhigh expression also reduced the ability of invasion of HeLa cells. Finally, GSTθincreased the susceptibility of HeLa cells and HCT116 cells toγ-ray irradiation by reducing DNA damage repair protein, such as XRCC4, DNA ligase IV和Ku70.The present results, for the first time, demonstrated that GSTθplays an important role in growth, cell cycle progression, cell invasion and radiosensitivity in human uterine cervix cancer HeLa cells and human colon cancer HCT116 cells, which will provide pre-clinical evidence for GSTθas a new target in cancer therapy in near future.

Rudiments of GSTθ and Cancer Dependability

Abstract

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