title: Soluble CD40 ligands sensitize the epithelial ovarian cancer cells to cisplatin treatment.
abstract: OBJECTIVE
  CD154 (CD40L) is a protein that is primarily expressed on activated T cells and is a member of the TNF superfamily of molecules. It binds to CD40 on antigen-presenting cells (APC), which leads to many effects depending on the target cell type. Being an activator of immune cells, CD40L has also been shown to directly induce apoptosis in tumor cells by multiple mechanisms. To understand the role of sCD40L in regulating the proliferation of epithelial ovarian cancer cells treated or untreated with cisplatin.
  
  
  METHODS
  Epithelial ovarian cancer cells: SKOV3 and its cisplatin-resistant strain SKOV3/DDP cells were used to test the effect of sCD40L and cisplatin. The proliferation of SKOV3 and SKOV3/DDP cells were measured by MTT. Cell cycle was assessed by flow cytometry. The mRNA expressions of targeted genes were detected by qRT-PCR. The protein expressions were detected by Western blotting.
  
  
  RESULTS
  sCD40L showed a significant dose-dependence inhibitory effect on the proliferation of ovarian cancer cell lines. sCD40L in combination with cisplatin could sensitized SKOV3/DDP cells to cisplatin treatment and reversed the drug resistance of SKOV3/DDP cells. The reversal ratios of 1 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 2.11, 2.71, while the reversal ratios of 2 μg/ml sCD40L combined with cisplatin in SKOV3 and SKOV3/DDP cells were 3.78, 5.20, respectively. sCD40L or sCD40L combined cisplatin increased tumor cells in G0/G1 phase. sCD40L in combination with cisplatin decreased the expression levels of GST-π, LRP, Survivin, p53 and Bcl-2 in both epithelial ovarian cancer cell lines. The protein expression level of GST-π, LRP and P53 protein was also decreased upon sCD40L in combination with cisplatin although the expression level of Bcl-2 and survivin protein had no significant difference.
  
  
  CONCLUSIONS
  sCD40L inhibits the proliferation of SKOV3 and SKOV3/DDP cells. The combined application of sCD40L and cisplatin can strength the inhibitory effect of cisplatin, and to a certain extent, reversing the resistance to cisplatin in SKOV3/DDP cells. sCD40L could lead a cell block in G0/G1 phase and make the cell growth restrained. sCD40L could induce SKOV3 and SKOV3/DDP cells apoptosis and reverse drug resistance through cutting GST-π mRNA, LRP mRNA, survivin mRNA, p53 mRNA and Bcl-2 mRNA and decreasing the expression of GST-π, LRP and P53 protein in SKOV3 and SKOV3/DDP cells, which provides in-vivo experiment basis to the application of sCD40L as a drug improving ovarian cancer cells sensitivity to cisplatin.
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Lijun
authors.initials: L
authors.lastname: Qin
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China; Department of Gynecology and Obstetrics, XingTai People's Hospital Affiliated to Hebei Medical University, Xingtai 054000, China.
authors.forename: Hongbing
authors.initials: H
authors.lastname: Qiu
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China; Department of Gynecology and Obstetrics, Baoding Hospital for Women and Children's Health Care, Baoding 071000, China.
authors.forename: Minjie
authors.initials: M
authors.lastname: Zhang
authors.affiliation: Department of General Surgery, Hebei General Hospital, Shijiazhuang 050000, China.
authors.forename: Fenghua
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Hongfang
authors.initials: H
authors.lastname: Yang
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Liu
authors.initials: L
authors.lastname: Yang
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Li
authors.initials: L
authors.lastname: Jia
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Kaiyun
authors.initials: K
authors.lastname: Qin
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Ling
authors.initials: L
authors.lastname: Jia
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Xiaomeng
authors.initials: X
authors.lastname: Dou
authors.affiliation: Department of Gynecology and Obstetrics, Zhaoxian County Hospital of Traditional Chinese Medicine, Shijiazhuang, 051530, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Cheng
authors.affiliation: Research Center, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Meixiang
authors.initials: M
authors.lastname: Sang
authors.affiliation: Research Center, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China.
authors.forename: Chao
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Gynecology and Obstetrics, Zhaoxian County Hospital of Traditional Chinese Medicine, Shijiazhuang, 051530, China.
authors.forename: Baoen
authors.initials: B
authors.lastname: Shan
authors.affiliation: Department of Gynecology, Fourth Hospital of Hebei Medical University, Shijiazhuang 050011, China. Electronic address: zzm701105@hotmail.com.
authors.forename: Zhengmao
authors.initials: Z
authors.lastname: Zhang
mesh_terms: D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D019013:CD40 Antigens / Q000235:genetics / Q000378:metabolism; D023201:CD40 Ligand / Q000494:pharmacology*; D000077216:Carcinoma, Ovarian Epithelial; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D002945:Cisplatin / Q000494:pharmacology*; D005260:Female; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism; D009375:Neoplasms, Glandular and Epithelial / Q000235:genetics / Q000473:pathology*; D010051:Ovarian Neoplasms / Q000235:genetics / Q000473:pathology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012995:Solubility
chemical_list: D019013:CD40 Antigens; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D023201:CD40 Ligand; D002945:Cisplatin
keywords: Cell apoptosis; Multidrug resistance; Ovarian cancer cell; sCD40L