1.Screening for differential genes of the prostate cancer and bioinformatics analysis of their interaction
Qianlin XIA ; Menglin SHAN ; Tao DING ; Yanjun ZHU ; Jun HOU ; Jianghua ZHENG
China Oncology 2017;27(3):169-176
Background and purpose: Gene chip is a nucleic acid sequence analysis method which is based on hybridization. It is a high-through put assay which can widely detect the level of gene expression in different tissues and cell types. This study aimed to compare and bioinformatically analyze differentially expressed genes between higher malignant degree of prostate cancer tissues and prostate inflammation tissues. Methods: The total RNAs were isolated from tissues of prostate cancer and prostate inflammation by TRIzol method and then purified, reversely tran-scribed to cDNA with incorporating biotin labeling probe, hybridized with Affymetrix Human U133 Plus 2.0 (covering 47000 transcripts,representing 38500 distinct genes). Picture signals of fluorescence in gene array were scanned and differential expression of gene in two tissues were compared by Command Console Software 4.0. These differential expressed genes were analyzed by bioinformatics methods finally. Results: According to the fold change ≥2, P<0.05, 1819 differential expression genes including 1025 up-regulated genes and 794 down-regulated genes were discovered. GO enrichment analysis displayed that these differentially expressed genes were mainly involved in cell cycle, cell metabolism, etc. KEGG pathway analysis found that these genes were mainly involved in some metabolism pathways including purine nucleotide metabolism. The interactions between the proteins encoded by these genes were analyzed by STING. Twenty key nodes genes including TPX2, ANLN, NUSAP1, MELK, DLGAP5, KIF11, TOP2A, RRM2 were dis-covered. Then this study revealed CEP55 and ANLN might be related to the occurrence and metastasis of prostate cancer by looking through literature. Conclusion: During the development of prostate cancer, the activation of genes related to cell cycle and cell migration, the abnormalities of genes related to metabolism and the inhibition of genes related to cell adhesion play critical roles in the development of prostate cancer. CEP55 and ANLN were related to the occurrence and prognosis of prostate cancer by systematic analysis which provided a valuable clue for the next experiment.
2.Detection ofHIC1 promoter methylation in prostate cancer using MSP and BSP methods
Menglin SHAN ; Tao DING ; Jianghua ZHENG ; Zenghui CHENG ; Ying GUO ; Qianlin XIA
China Oncology 2016;26(4):290-296
Background and purpose:Hypermethylated in cancer 1 (HIC1) is silenced in multiple cancer cells and tissues by DNA methylation of epigenetic modification, which may modulate the initiation and progression of tumors. However, there are few reports about this phenomenon in prostate cancer. This study aimed to investigate the status of HIC1 promoter methylation in prostate cancer using methylation methods.Methods:Methylation-specific polymerase chain reaction (MSP) and bisulfate sequencing PCR (BSP) were used to detect the methylation status ofHIC1 promoter in prostate cancer cell lines PC3 and C4-2B, prostate normal cell line PrEC, primary Chinese PCa tissues and the respective healthy control cases.HIC1 expression level was respectively determined by reverse transcription-PCR (RT-PCR) and Western blot assays in PC3, C4-2B and PrEC cells treated with 5-Aza-CdR.Results:We found that the percentages of HIC1 promoter methylation were 78.23%, 72.15% and 10.63% in PC3, C4-2B and PrEC cells by MSP analyses. Moreover, the levels of methylatedHIC1 promoter in 36 primary Chinese PCa tissues compared with the respective healthy control cases were 80.30%vs 31.56%. Expressions ofHIC1 mRNA and protein level were restored in PC3 and C4-2B cells after 5-Aza-CdR treatment.Conclusion:These findings demonstrate thatHIC1 promoter region is hypermethylated in prostate cancer, which results in silence or downregulation ofHIC1. The status ofHIC1 methylation can be a valuable marker in the early stage of prostate cancer and a potential therapeutic target.
3.Quantitative determination of phenolic characteristic components and their quality representation and correlation analysis of specific chromato-grams in Shaji
Yuanyuan ZHANG ; Shan LYU ; Mengjie CHEN ; Jing KONG ; Rendi SUN ; Yuanyuan ZHANG ; Dong-Bo LU ; Qiuhong WU ; Wenxia LI ; Menglin WANG ; Yuanyuan FAN ; Xin FENG ; Sitong LI ; Yu-Juan HUANG ; Yanyan JIANG ; Renbing SHI
Journal of Beijing University of Traditional Chinese Medicine 2018;41(5):383-394
Objective To establish an analysis method for quantitatively determining phenolic character-istic components and correlation analyzing their quality representation of specific chromatograms in Shaji (Seabuckthorn Fruit,Hippophae rhamnoides),and to review the quality of Shaji effectively and accurate-ly by applying association analysis-reviewing mode.Methods HPLC-PDA method was used to quantita-tively determine the content of phenolic characteristic components in 11 batches of Shaji(protocatechuic acid,ellagic acid,narcissin,quercetin,isorhamnetin),and to establish phenolic specific chromatograms of Shaji.The quality of 11 batches of Shaji was characterized based on teasing characteristic peaks and chemical types.The quantity of 11 batches of Shaji was characterized based on quantity and peak areas of protocatechuic acid, ellagic acid, narcissin, quercetin, isorhamnetin, phenolic acids(represented by protocatechuic acid)and flavonoids(represented by narcissin)in the specific chromatograms.The characterized results of quality and quantity of 11 batches of Shaji were given association analysis based on baseline material of Shaji.Results The characteristic components of protocatechuic acid, ellagic acid,narcissin,quercetin,isorhamnetin all had good linear correlation,and the results of methodological investigation were in accordance with the quantitative determination requirements.Taken batch 7 of Shaji as baseline material, there were totally 18 characteristic peaks in phenolic specific chromatograms of Shaji,including 3 peaks of phenolic acids and 15 peaks of flavonoids, and all 18 characteristic peaks appeared in the chromatograms of 11 batches of Shaji.The quantity of characteristic components were higher in batch 8, 7, 10, 4, 5, 11 and 1 after analyzed and reviewed by using association analysis-reviewing mode.The relevance of batch 6, 2, 3, 1 and 11 was the highest with baseline material of Shaji.Comprehensive reviewing showed that the excellent extents of batch 1,11,8,6 and 7 were prior. Conclusion The quantitative determination method of phenolic characteristic components in Shaji estab-lished in this study is easy and accurate.The association analysis-reviewing mode for quality characteriza-tion of phenolic specific chromatograms can be used for analyzing the quality and application validity of Shaji and reviewing quality of Shaji effectively and accurately.