title: [Effect of jianpi-jiedu formula on tumor angiogenesis-relevant genes expression in colorectal cancer].
abstract: To investigate the effect of the jianpi-jiedu formula (JPJD) on the expression of angiogenesis-relevant genes in colon cancer.  Methods: Crude extract was obtained from JPJD by water extract method. The effect of JPJD crude extract on colon cancer cell proliferation capacity was determined by MTT assays. The IC50 value was calculated by GraphPad Prism5 software. Affymetrix gene expression profiling chip was used to detect significant differences in expressions of genes after JPJD intervention, and pathway enrichment analysis was performed to analyze the differentially expressed genes. Ingenuity Pathway Analysis software was applied to analyze differentially expressed genes relevant to tumor angiogenesis based on mammalian target of rapamycin (mTOR) signaling pathway and then the network diagram was built. Western blot was used to verify the protein levels of key genes related to tumor angiogenesis.  Results: JPJD crud extract inhibited the proliferation capacity in colon cancer cells. The IC50 values in 24, 48, and 72 hours after treatment were 13.060, 9.646 and 8.448 mg/mL, respectively. The results of chip showed that 218 genes significantly upgraded, and 252 genes significantly downgraded after JPJD treatment. Most of the genes were related to the function of biosynthesis, metabolism, cell apoptosis, antigen extraction, angiogenesis and so on. There were 12 differentially expressed angiogenesis genes. IPA software analysis showed that the JPJD downregulated expression of sphingomyelin phosphodiesterase 3 (SMPD3), VEGF, vascular endothelial growth factor A (VEGFA), integrin subunit alpha 1 (ITGA1), cathepsin B (CTSB), and cathepsin S (CTSS) genes, while upregulated expressions of GAB2 and plasminogen activator, urokinase receptor (PLAUR) genes in the colorectal cancer cell. Western blot results demonstrated that JPJD obviously downregulated expressions of phospho-mTOR (P-mTOR), signal transducer and activator of transcription 3 (STAT3), hypoxia inducible factor-1α (HIF-1α), and VEGF proteins, while obviously upregulated the level of phospho-P53 (P-P53) protein.  Conclusion: JPJD may inhibit colorectal tumor angiogenesis through regulation of the mTOR-HIF-1α-VEGF signal pathway.
authors.affiliation: Department of Integrated Chinese and Western Medicine, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Dan
authors.initials: D
authors.lastname: Mao
authors.affiliation: Department of General Surgery, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Sanlin
authors.initials: S
authors.lastname: Lei
authors.affiliation: Department of Oncology, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Jin'an
authors.initials: J
authors.lastname: Ma
authors.affiliation: Department of Integrated Chinese and Western Medicine, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Li
authors.initials: L
authors.lastname: Shi
authors.affiliation: Department of Integrated Chinese and Western Medicine, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Shaofan
authors.initials: S
authors.lastname: Zhang
authors.affiliation: Hunan University of Traditional Chinese Medicine College of Pharmacy, Changsha 410028, China.
authors.forename: Jianhua
authors.initials: J
authors.lastname: Huang
authors.affiliation: Department of Pharmacy, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Xinyi
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Pharmacy, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Dengfeng
authors.initials: D
authors.lastname: Ding
authors.affiliation: Department of Integrated Chinese and Western Medicine, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Yingjin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Graduate School, Hunan University of Traditional Chinese Medicine, Changsha 410028, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Feng
authors.affiliation: Department of Integrated Chinese and Western Medicine, Second Xiangya Hospital, Central South University, Changsha 410011, China.
authors.forename: Sifang
authors.initials: S
authors.lastname: Zhang
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002401:Cathepsin B / Q000187:drug effects / Q000378:metabolism; D002403:Cathepsins / Q000187:drug effects / Q000378:metabolism; D045744:Cell Line, Tumor / Q000187:drug effects*; D015179:Colorectal Neoplasms / Q000098:blood supply / Q000235:genetics*; D015536:Down-Regulation; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000187:drug effects / Q000378:metabolism; D039001:Integrin alpha Chains / Q000187:drug effects / Q000378:metabolism; D009389:Neovascularization, Pathologic / Q000235:genetics; D055293:Receptors, Urokinase Plasminogen Activator / Q000187:drug effects / Q000378:metabolism; D050796:STAT3 Transcription Factor / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction; D013108:Sphingomyelin Phosphodiesterase / Q000187:drug effects / Q000378:metabolism; D058570:TOR Serine-Threonine Kinases / Q000187:drug effects / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000378:metabolism; D015854:Up-Regulation; D042461:Vascular Endothelial Growth Factor A / Q000187:drug effects / Q000378:metabolism
chemical_list: D004365:Drugs, Chinese Herbal; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D039001:Integrin alpha Chains; D055293:Receptors, Urokinase Plasminogen Activator; D050796:STAT3 Transcription Factor; D016159:Tumor Suppressor Protein p53; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C546842:MTOR protein, human; D058570:TOR Serine-Threonine Kinases; D013108:Sphingomyelin Phosphodiesterase; D002403:Cathepsins; D002401:Cathepsin B; C028217:cathepsin S