| Background and Objective:Gliomas account for about 46% of intracranial tumors, and represents one of the most aggressive and lethal human cancer types. Over the past decades, an increasing amount of evidences have showed that the existence of glioma stem cells. Glioma stem cells are a small part of glioma cells,it has lead to tumorigenesis, sustain and maintain ability of tumor heterogeneity. Cancer stem cells can be produce the new cancer cells, the root of tumor recurrence and metastasis. If the traditional treatment ignore this part of the cell, even though most of the other cancer cells are destroyed, the tumor will still be faced with the possibility of recurrence, and more resistant to treatment. Therefore, to kill cancer stem cells will be to determine the long-term efficacy of an important part of the tumor, unfortunately, all of the radiotherapy and chemotherapy were unable to kill tumor stem cells.Tumorigenesis is a complex process, both abnormal activation of oncogenes and inactivation of tumor suppressor genes there. PTEN genes involved in regulation of tumor stem cell signaling pathways. Therefore, we hypothesized that PTEN gene may by acting on glioma stem cells, inhibit the drying characteristics of glioma stem cells, thus inhibiting the development of glioma.In the present study, we not only want to explore stem characteristics of glioma stem cells and expression of PTEN in glioma tissue and glioma stem cells, we also constructed by PTEN lentivirus explore PTEN gene regulate Stem characteristics, invasion, apoptosis in glioma stem cell U87 s.Materials and Methods:1.Cultured glioma stem cells and detect the stem characteristics of U87 s.2. Detection of PTEN gene and protein expression in glioma tissues and normal brain tissue.3. PTEN gene lentiviral constructed and transfected glioma stem cells and determination of cell transfection efficiency.4. Effect on glioma stem cell U87 S through PTEN gene.5. PTEN gene affect the ability glioma stem cell invasion and apoptosis.6. Statistical analyses were performed using SPSS version 13.0, and significance determined with two-tailed Student’s t test. Data were considered statistically significant at p <0.05.Result:1. Culture and identification of glioma stem cells U87SIn order to study glioma stem cells, we used reverse culture from U87 glioma cell lines obtained glioma stem cells. By immunofluorescence to detect the expression of glioma stem cells U87 S markers CD133,A2B5, ALDH1,Nestein,, PLAGL2.The result showed that the glioma stem cells not only high expression of stem cell markers, but also has more ability to differentiate, self-renewal capacity. The above data shows that our cultured glioma stem cells to meet the characteristics of stem cells, the next step to meet the requirements of the experiment.2. The expression of PTEN gene and proteinWe found that PTEN gene expression in the glioma tissues and glioma stem cells U87 S significantly lower than the normal brain tissue. The experimental basis for our next experiment.3. PTEN gene affect glioma stem cells U87 S features of stem.We found that PTEN gene transfected glioma stem cell markers(CD133, A2B5, Nestein, SOX2, ALDH1, PLAGL2) expression was significantly decreased self-renewal rate is significantly lower than control group. PTEN gene may inhibit glioma stem cell characteristics, thereby inhibiting the development of glioma.4. PTEN gene affect the ability glioma stem cell invasion and apoptosis.We found that PTEN gene can not only inhibit the invasion ability of glioma stem cells and reduce the invasion protein MMP2, MMP9 expression, but also induce early apoptosis of glioma stem cells.Conclusions:In the present study, we demonstrated that PTEN as a tumor suppressor gene, by inhibiting stem glioma stem cell characteristics, invasion ability and promote apoptosis of glioma stem cells, thereby inhibiting the progression of gliomas. The result provide new insights into the development of therapeutic strategies against cancer via PTEN. |
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