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SIRNA interfere with transcription factor SOX2 and OCT4 on the proliferation of cancer stem cells

Author: HuZuo
Tutor: HuXiang
School: Dalian Medical University
Course: Department of General Surgery
Keywords: Gastric Cancer Transcription factor SOX2 Transcription factor 0CT4 cancer stem cells 5-FU
CLC: R735.2
Type: Master's thesis
Year: 2011
Downloads: 89
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Abstract


Background: surgery, chemotherapy, radiotherapy and other traditional means of advanced gastric cancer is difficult to curb the recurrence, metastasis, and the presence of cancer stem cells may be the recurrence and metastasis of gastric cancer is one of the main factors. There is no targeted treatment of cancer stem cells. Known transcription factors OCT4 and SOX2 in cancer stem cell formation and maintenance of cancer stem cell properties and function play an important role, so the interference of these genes inhibit tumor stem cells affect worth exploring. Objective: To investigate the interference SOX2 and OCT4 gene on human gastric cancer stem cell proliferation. Propose a new perspective for the cancer stem cell therapy for the treatment of gastric cancer has opened new ways to lay the theoretical and experimental basis. Method: 1. Using RT-PCR method to detect human gastric carcinoma cells (BGC-823) transcription factor SOX2 and OCT4 expression. 2 chemotherapy treatment and enrichment of stem cells in gastric cancer cells. 3 by flow cytometry before and after chemotherapy cancer stem cell surface markers CD44 ratio. 4 Design, Synthesis siRNA-sox2 and siRNA-oct4, located siRNA-SOX2 group (transfected with siRNA-SOX2), siRNA-OCT4 group (transfected with siRNA-OCT4), co-transfection group (co-transfected siRNA-SOX2 and siRNA-OCT4) group and blank control group (only adding transfection reagent without siRNA) and normal control group and transfected chemotherapy drugs before treatment and after treatment of gastric cancer cells and enrichment of cancer stem cells. By RT-PCR, RNA interference at the mRNA level validation results. 5. Flow cytometry before and after treatment with chemotherapy drugs interfering RNA transcription factor SOX2 and OCT4 on gastric cancer cell cycle changes. 6.MTT measured before and after treatment with chemotherapy drugs interfering RNA transcription factor SOX2 and OCT4 on gastric cancer cell proliferation. Results: 1. Gastric cancer BGC-823 cells in the presence of transcription factors SOX2 and OCT4 expression. (2) chemotherapy drug 5-FU treatment of gastric cancer cells, the vast majority of cell death, survival was 7 ± 0.202%; cell cycle in G0/G1 phase cells was significantly higher than before treatment (89.76% ± 4.93% VS 68.19% ± 2.92%, P lt; 0.01), S phase cells compared with before treatment was significantly lower (0% VS 23.94% ± 1.98%, P lt; 0.01), viable cells of stem cell marker CD44 in proportion than before treatment was significantly higher (10.14 ± 0.192% VS 0.74 ± 0.052%, P lt; 0.01). 5-FU treatment transcription factor SOX2 expression was significantly higher than before treatment (25 ± 0.41% VS 8 ± 0.3%, P lt; 0.01); transcription factor OCT4 expression than before treatment was also significantly increased (83 ± 4.7% VS 20 ± 2.1%, P lt; 0.01). Description chemotherapy drug 5-FU treatment of residual cancer cells rich in cancer stem cells. 3.siRNA-sox2 and siRNA-oct4 transfected into gastric cancer cells, SOX2 expression increased from the pre-disturbance 8 ± 0.3% down to 1 ± 0.2%, P lt; 0.01. OCT4 expression increased from 25 ± 2.1% before interference decreased to 2 ± 0.4%. 4.siRNA interfere with cancer cell transcription factor after siRNA-SOX2 group, siRNA-OCT4 group of transfected group and blank group M/G2 phase cells were significantly lower (P lt; 0.01), with no significant interference group differences in the proportion of cells in S phase interference group were significantly higher than normal (P lt; 0.05), no significant difference between the interference group. 5.siRNA interfere with gastric cancer cell line BGC-823 transcription factors SOX2 and OCT4 after, siRNA-SOX2 group, siRNA-OCT4 group, co-transfection group and blank control group (only liposome transfection, non-specific siRNA) compared to its cell proliferation was significantly slowed down, the peak delay P lt; 0.01, each RNA interference was no statistical difference between the groups. After the application of siRNA chemotherapy drug 5-FU, interference in each group compared with the control group, the sensitivity of cells to chemotherapeutic drugs significantly increased (P lt; O.O1), no difference in the interference group. siRNA interference with 5-FU treatment of gastric cancer cells siRNA-SOX2 group, siRNA-OCT4 group, co-transfection group compared with the control group, respectively, significantly inhibited proliferation, P lt; 0.01. Conclusions: 1. Poorly differentiated human gastric cancer cell line BGC-823 expression in stem cell-related transcription factors SOX2 and OCT4. 2. Chemotherapy treatment after gastric cancer cells significantly increased the proportion of stem cells, cancer stem cells can achieve the purpose of enrichment. 3 prior chemotherapy treatment, RNA interference target gene can slow down the proliferation of gastric cancer cells, the peak latency, and so the sensitivity to 5-FU increased significantly. Chemotherapy treatment, RNA interference can significantly inhibit tumor stem cell enriched gastric cancer cell proliferation, rather than continuous tumor effect of chemotherapy drugs given to more significant. Tips chemotherapy drug combination therapy with RNA interference can enhance the efficacy of anti-cancer therapy. Objective transcription factor SOX2 and OCT4 in maintaining cancer stem cells play an important role in the process, is the future of cancer stem cell therapy is one of the potential targets, through RNA interference interference stem cell related genes may become a new strategy for the future treatment of cancer.

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CLC: > Medicine, health > Oncology > Gastrointestinal Cancer > Gastric neoplasms
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