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Combined Effects of klotho and Soluble CD40 Ligand on A549 Lung Cancer Cells

Author: WangXueLi
Tutor: WuJianQing
School: Nanjing Medical University
Course: Geriatrics
Keywords: Lung cancer Klotho sCD40L Apoptosis
CLC: R734.2
Type: Master's thesis
Year: 2011
Downloads: 25
Quote: 0
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Abstract


BACKGROUND: Lung cancer is one of the common malignant tumors. In many countries, the incidence of lung cancer ranks first in the malignant tumor, only about 10% of the five-year survival rate in patients with non-small cell lung cancer. While traditional treatment has made great progress, but has not been significantly improved overall survival of lung cancer patients. Therefore important to explore new treatment. At present, more and more studies have shown that the gene therapy of tumors is an effective method. Klotho gene is a new type of anti-aging gene, can effectively increase the immune function, but now more and more studies show that the Klotho gene is closely associated with cancer. CD40 ligand (CD40L) is one of the TNF superfamily, first found expression in activated CD4 ~ T lymphocyte surface, have an important role in the cellular immunity and humoral immunity has been demonstrated for a variety of malignant cells CD40 expression on the direct and indirect anti-tumor effect of CD40 excited. Since sCD40L and Klotho gene both tumor and immune role, we speculated that the combination of the two can further inhibit tumor growth and promote tumor cell apoptosis. Objective: This study was designed to investigate the biological role of Klotho gene soluble CD40 ligand of A549 lung cancer cells (CD40 positive expression cell lines), and its related mechanism. Methods: Quantitative RT-PCR was observed Klotho expression plasmid pCMV6-MYC-KL Klotho mRNA expression levels in the transfected A549 cells; MTT method was used to observe cell growth activity changes and growth curve; flow cytometry to detect apoptosis Quantitative RT-PCR to detect the expression of apoptosis-related genes Bax/Bcl-2 change. The results: (1) the Klotho gene Joint sCD40L than individually transfected the Klotho gene or join sCD40L increased growth inhibition of cell A549 (p <0.05). (2) flow cytometry showed that the Klotho gene combined of sCD40L role in cell A549, apoptosis rate than separate transfection Klotho or added separately sCD40L increased (p <0.05). (3) Klotho gene Joint sCD40L role in A549 cells 48 hours after the decline of the Bcl-2 gene expression than individually transfected Klotho gene or join sCD40L alone increased (p <0.05), separate Klotho gene transfection allows Bax gene expression The amount raised (p <0.05), but added separately the sCD40L groups Bax gene expression than blank control group had no significant difference (p ≧ 0.05). Conclusion: Klotho gene and sCD40L alone and the combination of the two may be to inhibit the proliferation of lung cancer A549 cells raised the Bax gene expression and / or down-regulated expression of Bcl-2 gene expression by inhibiting cell cycle, and induce apoptosis; their combination after A549 cells above is further enhanced.

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CLC: > Medicine, health > Oncology > Respiratory system tumors > Lung tumors
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