title: Hydroxycamptothecin-induced apoptotic gene expression profiling by PCR array in human Tenon's capsule fibroblasts.
abstract: Studies have indicated that hydroxycamptothecin (HCPT) induces apoptosis of fibroblasts. In this study, we investigated the apoptotic gene expression profiling in HCPT-treated -human Tenon's capsule fibroblasts (HTCFs) and identify the most implicated gene in apoptotic signaling of HTCFs by HCPT. Method HTCFs were incubated with HCPT at 0, 0.25 and 4 mg/L for 24, 48 and 72 h, respectively. Anti-proliferative effects were measured by MTT assay. Apoptosis was determined using the Annexin V-FITC/PI apoptosis detection kit and apoptotic cells were identified by flow cytometry. A PCR array was employed to analyze the most implicated apoptotic genes during HCPT-induced apoptosis in HTCFs. Results from our studies showed that HCPT induced apoptosis in HTCFs in a concentration-and time-dependent manner. The apoptotic HTCFs was increased by 9.38% with a multiplicity at 4 mg/L HCPT. PCR array demonstrated remarkable changes in 88 apoptotic genes, including 9 up-regulated genes and 36 down-regulated genes. HCPT treatment induced the upregulation of CHOP and downregulation of XIAP in HTCFs. To conclude, our results support HCPT induced the apoptosis of HTCFs, involving the activation of mitochondrial and endoplasmic reticulum stresses as well as the downregulation expression of XIAP.
authors.affiliation: Department of Ophthalmology, Wuxi No. 2 People's Hospital Affiliated Nanjing Medical University Wuxi 214002, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Tang
authors.affiliation: Department of Ophthalmology, Wuxi No. 2 People's Hospital Affiliated Nanjing Medical University Wuxi 214002, China.
authors.forename: Yinong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology, Wuxi No. 2 People's Hospital Affiliated Nanjing Medical University Wuxi 214002, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhang
authors.affiliation: Department of Ophthalmology, Wuxi No. 2 People's Hospital Affiliated Nanjing Medical University Wuxi 214002, China.
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Ophthalmology, Wuxi No. 2 People's Hospital Affiliated Nanjing Medical University Wuxi 214002, China.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Wu
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D002166:Camptothecin / Q000494:pharmacology*; D002478:Cells, Cultured; D059865:Endoplasmic Reticulum Stress / Q000187:drug effects; D005347:Fibroblasts / Q000187:drug effects*; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D060888:Real-Time Polymerase Chain Reaction; D058475:Tenon Capsule / Q000187:drug effects*; D059467:Transcriptome / Q000187:drug effects*; D051636:X-Linked Inhibitor of Apoptosis Protein / Q000096:biosynthesis
chemical_list: D051636:X-Linked Inhibitor of Apoptosis Protein; C496866:XIAP protein, human; D002166:Camptothecin
keywords: HCPT; HTCFs; XIAP; signal pathway