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Inhibition of survivin expression and mechanisms of reversing drug-resistance of human lung adenocarcinoma cells by siRNA

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Chinese Medical Journal 2010年第20期123卷 2901-2907页

作者： LIU Jing-lei WANG Yan JIANG Ji KONG Rui YANG Yan-mei JI Hong-fei SHI Yu-zhi Department of Medical Oncology Affiliated Tumor Hospital of Harbin Medical UniversityHarbinHeilongjiang 150081China Department of Isotope Affiliated Tumor Hospital of Harbin Medical UniversityHarbinHeilongjiang 150081China Department of Surgery First Affiliated Hospital of Harbin Medical UniversityHarbinHeilongjiang 150001China Harbin Medical University Tumor Institution HarbinHeilongjiang 150081China Department of Respiratory Medicine First Affiliated Hospital of Harbin Medical UniversityHarbinHeilongjiang 150001China

Background Survivin, a member of the inhibitor of apoptosis protein （IAP） family, overexpresses in tumor cells and not expresses in terminally differentiated adult tissues. This study aimed to investigate the effects of survivin-specific siRNA on cell proliferation, apoptosis and chemosensitivity to cisplatin in vitro and in vivo and explore the mechanisms about decreasing expression of survivin in reversing cancer cells resistance to chemotherapeutic *** Survivin-specific siRNA was transfected into A549/DDP cells. The expression of survivin and lung resistance-related protein （LRP） mRNA levels were determined by RT-PCR, chemosensitivity of A549/DDP （cisplatin）cells to cisplatin was determined by MTT assay, and apoptosis and cell cycle were determined by flow cytometry （FCM）.The protein expression levels of survivin, LRP, cyclin-D1, caspase-3 and bcl-2 were determined by Western blotting analyses. The effect of survivin siRNA inhibition on tumor growth was studied in athymic nude mice in *** Survivin-specific siRNA efficiently down-regulated survivin expression. The cell cycle was arrested at G2/M phase, and apoptosis was obviously found. Inhibition of survivin expression could make the IC50 and drug-resistant index of cisplatin decrease, and enhance the cancer cells sensitivity to cisplatin. After transfection by survivin-specific siRNA, expression of LRP and cyclin-D1 were downregulated, caspase-3 expression was upregulated, bcl-2 expression had no obvious change. The animal experiment confirmed knockdown of survivin could inhibit the tumor *** Survivin-specific siRNA can efficiently suppress the expression of survivin, increase apoptosis, inhibit cells proliferation and enhance the chemosensitivity to cisplatin in vitro and in vivo. Suppression of survivin expression helping to reverse drug-resistance may have relationship with downregulation of LRP and upregulation of ***-tumor strategies based on the inhibition of sur

关键词： human lung adenocarcinoma survivin chemosensitivity siRNA apoptosis

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The Study on the Activity against lung adenocarcinoma A549 by Combination of human γδ T Cells with Small Molecule Compound

The Study on the Activity against Lung Adenocarcinoma A549 b...

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浙江省免疫学会第十次学术大会

作者： Li-Li Ma Xing-jian Cheng Yan-Ling Wu Yoshimasa Tanaka Wen Zhang Department of Biopharmaceutical Sciences College of Pharmaceutical SciencesZhejiang University of Technology Lab of Molecular Immunology of Zhejiang Provincial Center for Disease Control and Prevention

Objective:In order to research the therapeutic effects of yST cells or γδT cells combining other small molecule compound on lung adenocarcinoma cancer A549 in nude mice and their antitumor ***:Firstly,Zoledronic acid(Zol) combined EL-2 were used to augment γδT cells from peripheral blood mononuclear cells(PBMCs).The purity of expanded γδT cells were assessed by flow ***,the ability of yST cells or γδT cells combined small molecule compound inhibiting A549 cells in vitro wasmonitored by utilizing Real-Time Cell Analysis(RTCA) ***,animal model of A549 in Balb/c nude mice was established to explore the anti-tumor effect of γδ T cells alone or co-administration withcompound and ***,the expression of mRNA and protein of c-Myc and PD-1 in the tumor tissue were measured by Reverse Transcription Polymerase Chain Reaction(RT-PCR) and ***:Flow cytometry results showed that γδT cells have high purity after co stimulating by Zo1 and IL-2 for 14 ***-Time Cell Analysis demonstrated that γδT cells or γδT cells combined compound could inhibit A549 cells growth,and γδT cells combined compound had stronger inhibitory *** the observation of the volume and weight of nude mice,we found that:(1) Compared with healthy nude mice,nude mouse bearing the tumor gain less weight(p<0.05),that indicated tumor transplantation affects the normal physiological function of nude mice.(2) During therapy,the mice of γδT cells treatment group and combining treatment group had a smaller volume growth of tumor comparing to the Control group(p<0.05).Furthermore,the mass of the mice of γδT cells treatment group and combining treatment group is less than the mice of Control group.(3) RT-PCR and Western-blotting experiment results showed that the transcription and expression level of c-Myc and PD-1 in tumor tissue of γδT cells treatment group and combining treatment group were lower than control ***:γδT cells

关键词： γδT cells human lung adenocarcinoma immunotherapy

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