›› 2012, Vol. 32 ›› Issue (7): 891-.doi: 10.3969/j.issn.1674-8115.2012.07.015

• Original article (Basic research) • Previous Articles     Next Articles

Effects of UbcH10 gene silencing combined with paclitaxel treatment on proliferation and apoptosis of NCI-H226 cells

ZHAO Li-ming1, WANG Liang-zhe2, LOU Li-rong3, SUN Guang-yuan4, FANG Zheng1, XIU Qing-yu1   

  1. 1.Department of Respiratory Medicine, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China；2.Department of Pathology, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China；3.Shanghai Pudong New Area Health Inspection, Shanghai 200136, China；4.Department of Thoracic Surgery, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China
  • Online:2012-07-28 Published:2012-08-17
  • Supported by:

    Shanghai Municipal Health Bureau Foundation, 20090122

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Abstract:

Objective To investigate the effects of small interfering RNA(siRNA)-mediated UbcH10 gene silencing combined with paclitaxel treatment on proliferation and apoptosis of human lung squamous carcinoma cell line NCI-H226. Methods siRNA-UbcH10 sequence targeting UbcH10 gene was synthesized chemically, and was transfected into NCI-H226 cells by lipofectin (siRNA transfection group). Twenty-four hours after transfection, the expression of UbcH10 mRNA and protein was detected by Real-Time PCR and Western blotting. Cells without transfection were served as blank controls, and those transfected with negative sequence as negative controls. NCI-H226 cells transfected with or without siRNA were treated with paclitaxel (1 μmol/L)(siRNA+ paclitaxel group or paclitaxel group), cell proliferation was detected by MTT assay 24 h and 48 h after transfection, and cell apoptosis was determined by flow cytometry 24 h after transfection. NCI-H226 cells without paclitaxel treatment while with siRNA transfection, negative sequence transfection and no transfection were served as siRNA transfection group, negative control group and blank control group respectively. Results Real-Time PCR and Western blotting revealed that the expression of UbcH10 mRNA and protein in siRNA transfection group was significantly lower than that in blank control group and negative control group (P<0.01). MTT assay indicated that the cell proliferation inhibition rate in siRNA transfection group was significantly higher than those in paclitaxel group and control group (P<0.05), while the cell proliferation inhibition rate in siRNA+paclitaxel group was significantly higher than that in siRNA transfection group (P<0.05). The cell apoptosis rate in siRNA transfection group was significantly higher than those in paclitaxel group and control group (P<0.05), and the cell apoptosis rate in siRNA+paclitaxel group was significantly higher than that in siRNA transfection group (P<0.05), while there was no significant difference in the cell apoptosis rate between paclitaxel group and control group (P＞0.05). Conclusion UbcH10 gene silencing can significantly increase the sensitivity of NCI-H226 cells to the chemotherapy drug paclitaxel.

Key words: UbcH10 gene silencing, small interfering RNA, paclitaxel, NCI-H226 cell, cell proliferation, cell apoptosis