HPLC测定人肝转基因细胞中的盐酸维拉帕米

目的 :为了研究盐酸维拉帕米经人肝 CYP4 5 0转基因细胞的代谢 ,建立了反相高效液相色谱的测定方法。方法 :与人肝转基因细胞提取的 S9上清液孵育之后的样品用二氯甲烷 -异丙醇 ( 9∶ 1,V/V)提取 ,采用地西泮作为内标 ,以 Shim- packCL C- ODS C1 8为色谱柱 ,甲醇 -水 -三乙胺 ( 60∶ 4 0∶ 0 .4 ,V/V) ,p H6.4为流动相 ,紫外检测波长为 2 3 5 nm,流速 0 .5 m l·min- 1 。结果 :维拉帕米浓度在 5～ 3 2 0μg· ml- 1 范围内线性关系良好 ,r=0 .9999,n=6。检测限为 6.2 5 ng( S/N≥ 3 ) ,定量限为9.4 ng± 1.0 6ng,相对标准偏差 ( RSD) <12 % ( n=5 )。方法回收率达 94 .4～ 99.6% ,日内、日间 RS D分别为 5 .9～ 2 .9% ,5 .2～ 3 .8% ( n=5 )。结论 :此方法简便、准确 ,可用于研究盐酸维拉帕米在人肝转基因细胞中的代谢。

转基因肝星状细胞定向诱导骨髓Thy-1^+β_2M^-细胞向肝细胞分化

体内移植研究表明,骨髓来源的干细胞具有向成熟的肝细胞分化的能力。然而,成体干细胞向肝细胞诱导分化的效率、分化细胞的功能状态、微环境对细胞分析的影响等均存在许多疑问。以高效、稳定表达肝细胞生长因子的肝星状细胞株CFSC/HGF作为饲养细胞,成功地将大鼠骨髓来源的Thy-1+β2M-细胞(bone marrow derived Thy-1+β2M- cells,BDTCs)诱导为肝细胞,经RT-PCR和免疫荧光化学检测证实,该条件下诱导后的BDTCs表达幼稚或成熟肝细胞特异性的基因,分化细胞具有成熟肝细胞特有的靛青绿摄取排泌、氨基代谢和白蛋白分泌的功能,提示肝细胞生长因子、肝脏非实质细胞及其产生的细胞外基质为骨髓干细胞向肝细胞分化提供了适宜的微环境。这为以骨髓干细胞为基础的再生医学在终末期肝病治疗中的应用提供了理论和技术上的支持。

雷公藤红素对转基因小鼠原代肝细胞HBV的抑制作用

目的:考察雷公藤红素对HBV转基因小鼠原代肝细胞的抗HBV作用。方法:改良的两步灌流法分离HBV转基因小鼠原代肝细胞;MTT法检测细胞毒性作用;药物作用24 h后取上清,分别应用ELISA法和荧光定量PCR法测定HBsAg和HBV-DNA。结果:雷公藤红素对原代肝细胞TC50为(6.49±0.8)μmol·L-1;雷公藤红素浓度低于0.8μmol·L-1时,对原代肝细胞没有明显毒性;在0.08～0.8μmol·L-1浓度范围内对HBsAg和HBV-DNA有显著抑制作用(P<0.05);随着雷公藤红素浓度的增加,对HBsAg和HBV-DNA的抑制作用增强。结论:雷公藤红素不仅能够抑制转基因小鼠原代肝细胞HBV-DNA的复制,而且可以有效抑制其HBsAg的表达。

扁塑藤素对HepG2.2.15细胞生长的影响及其对乙肝病毒的抑制作用

目的研究扁塑藤素在体外对肝癌细胞株HepG2. 2. 15细胞生长的影响及对乙肝病毒(HBV)的抑制作用。方法将不同浓度的扁塑藤素作用于转染HBV全基因组DNA的肝癌细胞株HepG2. 2. 15,作用24 h后收集上清,采用MTT法测定扁塑藤素对HepG2. 2. 15细胞生长的影响,通过形态学观察扁塑藤素对HepG2. 2. 15细胞生长的影响;采用改良的两步灌流法分离HBV转基因小鼠原代肝细胞;采用MTT法观察不同浓度扁塑藤素对原代肝细胞的毒性;应用ELISA法和PCR法测定乙肝表面抗原(HBsAg)和HBV-DNA含量。结果扁塑藤素可以影响HepG2. 2. 15细胞的生长,显微镜下亦可见被药物作用后的细胞脱落明显,脱落细胞悬浮于培养液中,形态变圆,折光性差;扁塑藤素对原代肝细胞的最大无毒浓度为2μmol·L^-1,最大无毒浓度以下,浓度为2、0. 2、0. 02μmol·L^-1的扁塑藤素可以有效抑制原代肝细胞HBsAg和HBV-DNA的表达,且随着扁塑藤素浓度的减低对HBsAg和HBV-DNA的抑制作用减弱。结论扁塑藤素可以影响HepG2. 2. 15细胞的生长;在HBV转基因小鼠原代肝细胞中,一定浓度的扁塑藤素可以抑制其HBsAg和HBV-DNA的表达。

Transfection of apoptosis related gene Fas ligand in human hepatocellular carcinoma cells and its significance in apoptosis

AIM: To evaluate the expression of apoptosis related gene Fas ligand (FasL) in human hepatocellular carcinoma (HCC) cells HepG2 and its significance in apoptosis. METHODS: Levels of soluble Fas ligand (sFasL) in a group of patients with hepatitis B virus (HBV)-induced chronichepatitis, HBV-positive liver cirrhosis and HCC were evaluated. In a further study, the recombinant eukaryotic expression plasmid pcDNA3.1hisB-FasL was transfected into HCC cells HepG2 by lipofection, and then soluble FasL was examined in the supernatant of culture cells by EIA, FasL expression in HepG2 cells was detected by immuohistochemistry. After being stained by annexin V and propidium iodine, cells were passed through a flow cytometer and examined by a fluorescence microscope and a laser scanning microscope.RESULTS: The sFasL levels were significantly lower in patients with HCC when compared to the patients with hepatitis or liver cirrhosis. In comparison with untransfected cells, the soluble FasL could be detected in the supernatant of transfected cells. FasL was expressed on the membranes and cytoplasm of transfected cells. The apoptotic cell rate was 36.30% in transfected cells, and was 11.53% in untransfected cells. Moreover, the different stage of apoptotic cells could be distinguished by annexin V and propidium iodine staining.CONCLUSION: Fas ligand is an apoptotic pathway of HCC cells.

Mechanism of metastasis by membrane type 1-matrix metalloproteinase in hepatocellular carcinoma

AIM To investigate the precise role of membrane type 1-matrix metalloproteinase （NTI-NNP） in hepatocellular carcinoma （HCC） metastasis. METHODS- Human HCC cells Hep3B with overexpression of MTT-MMP were established by stable transfection, and compared with control cells carrying the empty vector. Cells were examined in vivo for their differences in the metastatic ability of athymic nude mice, and analyzed in vito for their differences in invasion ability by invasion chamber coated with Matrigel, adhesion towards collagen I and migration through culture chamber. Cell proliferation and apoptosis in adherent and suspension status were evaluated by MTr and flow cytometry analysis. RESULTS We found that overexpression of MT1-MMP could increase intrahepatic metastasis in nude mice with orthotopic implantation of HCC cells （incidence of 100% [MT1-MMP transfectants] vs 40% [vector control transfectants], P〈0.05）. NT1-MMP could also enhance cell invasion through Natrigel （107.7 vs 39.3 cells/field, P〈0.001）, adhesion towards matrix （0.30 vs 0.12 absorbance unit at 540 nm, P〈0.001）, cell migration （89.3 vs 39.0 cells/field, P〈0.001）, and cell proliferation （24.3 vs 40.5 h/doubling, P〈0.001）. We also observed that NTI-NNP supported cell survival （71.4% vs 23.9%, P〈0.001） with reduced apoptosis （43.7% vs51.0%, P〈0.05） in an attachment-free environment. CONCLUSION： MT1-MMP overexpression could enhance metastasis. In addition to its active role in matrix degradation during tumor invasion, MT1-MMP enhances tumor cell survival upon challenge of detachment, which is important during metastasis when cells enter the circulation.

The promoter analysis of the human C17orf25 gene, a novel chromosome 17pl3.3 gene

The human C17orf25 gene (Accession No. AF177342) is one of thirteen genes cloned from a region displaying a high score of loss of heterozygosity within chromosome 17pl3.3 in human hepatocellular carcinoma in China[l]. To unveil the underlying mechanisms for the transcription regulation of this gene and understand its implication to the hepatocellular carcinogenesis, we looked into the relevant aspects by both bioinformatic and experimental executions. We found: 1, The abundant expression of the C17orf25 gene was evident in all the cell lines and tissue samples tested, showing little hepatoma-selectivity; 2, Its transcription starts at a single site, locating at -60 from the translation initiation codon; 3, A 58 bp fragment containing the transcription start, extending from -112 to -55, represents the minimal promoter; 4, The consensus sequence within this fragment recognized by SP1 contributes predominantly to the activity of the minimal promoter; 5, The bioinformatic analysis suggests that the C17orf25 gene may encode a protein in the family of the glyoxalase. Our data has provided some deep insight into both function and regulation of the C1 7orf25 gene in the context of the normal liver and hepatocellular carcinoma.

Inhibitory effect of adeno-associated virus-mediated gene transfer of human endostatin on hepatocellular carcinoma

AIM: To investigate the effect of adeno-associated virusmediated gene transfer of human endostatin on the growth of hepatocellular carcinoma (HCC).METHODS: HCC cell line Hep3B was infected with recombinantadeno-associated virus containing human endostatin gene (rAAV2-hEndo). The results of transfection were detected by RT-PCR and SDS-PAGE assay. MTT assay was used to observe the effects of supernatant of transfected cells on ECV304 cell proliferation. An animal model of HCC was established by injecting Hep3B cells subcutaneously into the back of nude mice. Intratumoral injection of rAAV2hEndo, empty virus and phosphate-buffered saline were given sequentially. Serum endostatin was determined byELISA, the inhibitory effect of endostatin on the growth of xenograft was assessed in 3 wk.RESULTS: The results of RT-PCR and SDS-PAGE assay confirmed that rAAV2-hEndo successfully transfected Hep3B cells, and endostatin was secreted from Hep3B cells to medium. The supernatant of transfected cells markedly inhibited the proliferation of ECV304 cells (P＜0.01). Intratumoral injection of rAAV2-hEndo (2×1010v.g.) led to a sustained serum endostatin level ofapproximately (86.71±5.19) ng/mL. The tumor volumeand microvessel density were less in rAAV2-hEndo group than in control groups (P＜0.01).CONCLUSION: Human endostatin can be stably expressed by adeno-associated virus-mediated gene transfer and effectively inhibit the growth of HCC.

Cloning and characterization of human IC53-2,a novel CDK5 activator binding protein

We have identified IC53-2, a human homologue of the rat C53 gene from a human placenta cDNA library (GeneBank Accession No. AF217982). IC53-2 can bind to the CDK5 activator p35 by in vitro association assay. IC53-2 is mapped to human chromosome 17q21.31. The IC53-2 transcript is highly expressed in kidney, liver, skeletal muscle and placenta. It is abundantly expressed in SMMC-7721, C-33A, 3AO, A431and MCF-7 cancer cell lines by RT-PCR assay. Stable transfection of IC53-2 cDNA into the hepatocellularcarcinoma SMMC-7721 cell remarkably stimulates its growth in vitro. The above results indicate thatIC53-2 is a novel human gene, which may be involved in the regulation of cell proliferation.

Intrahepatic and peripheral T-cell responses in genotype 1b hepatitis C virus-infected patients with persistently normal and elevated aminotransferase levels

AIM： To evaluate whether the cytokine responses in liver and serum differ in chronic hepatitis C patients with normal and high alanine aminotransferase （ALT） levels. METHODS： Thirty-three （16 with normal ALT level as group 1 and 17 with elevated ALT level as group 2） patients infected with genotype lb hepatitis C virus （HCV） were examined. Liver infiltrating lymphomononuclear cells （LILMCs） were isolated from liver biopsy by collagenase type 1 and stimulated with phytohemagglutinin and interleukin 2 （IL-2）. IL-10, IL-12, interferon gamma （IFN-γ） and tumor necrosis factor alpha （TNF-α were determined in serum and LILMCs by ELISA. RESULTS： Serum cytokine levels were similar in both groups （P〉0.05）. Stimulated IFN-γ and TNF-α levels in LILMCs were increased in both groups. IL-12 and IL-10 levels stimulated with IL-2 were higher in group 1 than in group 2 （P = 0.023）. Histological activity index （HAI） and stage had a negative correlation with TNF-α and IFN-γ levels in group 2. CONCLUSION： Increased T-helper type 2 （Th2） cytokine response may regress inflammatory and biochemical activity. Progression of histological abnormalities in persons with elevated ALT probably depends on insufficient Th2 cytokine response, which does not balance Th1 cytokine response.

Cytotoxicity Research of Recombinant Human Lactoferrin on Primary Hepatocyte and Nephrocyte Cell

[Objective] The aim was to study whether recombinant Human Lactoferrin has toxic effect on Primary Hepatocyte and Nephrocyte Cell of rat to provide reference for further safety evaluation.[Method] Recombinant Human Lactoferrin and its digested products were taken as tested compound,cow Lactoferrin was used for contrast.Primary Hepatocyte and Nephrocyte Cell of rat were cultured and IC50 values were tested by MTT,and cytotoxic dose-response relationship was tested.[Result]Target toxicity was not found from recombinant Human Lactoferrin on hepatocytes and nephrocytes,in accordance with sub-chronic toxicity test.[Conclusion] This study is of reference value for further safety evaluation of recombinant Human Lactoferrin and safety of evaluation method of GM food.

THE INFLUENCE OF HUMAN SINGLE CHAIN INTELEUKIN-12 GENE TRANSDUCTION ON THE BIOLOGICAL BEHAVIOR OF HEPATOMA 7721 CELLS

Objective. To investigate the anti- tumor effects of human single chain interleukin- 12 (hscIL- 12). Method. pcDNA/ hscIL- 12 recombinant was transfected into human hepatic carcinoma cells (7721 cells) by lipofectin method. The 7721/hscIL- 12 cells which secrete hscIL- 12 stably, were obtained via G418 selection, and in vitro the influence of hscIL- 12 gene transduction on the growth of tumor cells was evaluated by cell cycle analysis. In vivo, genetically engineered 7721 cells (7721/hscIL- 12, 7721/pcDNA) and parental cells were implanted into BALB/c nude mice,respectively. 7721/pcDNA and 7721/hscIL- 12 groups were divided into two sub- groups on day 8: one was administered with hPBL twice, 6 days at interval; the other was given equal volume of PBS. Mice were sacrificed on day 26, and spleens and tumors were taken out for histologic assay. Results. hscIL- 12 produced stably by 7721/hscIL- 12 cells had bioactivity, and it was proved by Western blot, immunocytochemistry, and in situ hybridization. In vitro, compared with 7721 and 7721/pcDNA, the 7721/hscIL- 12 grew much more slowly. FACS assay showed apparent G1 arrest of 7721/hscIL- 12 cells. In animal experiment, on day 8 after inoculation, the tumors of 7721 and 7721/pcDNA group were up to 5～ 7mm,while those of 7721/hscIL- 12 group were 2～ 4mm.When treated with hPBL, the tumor of 7721/hscIL- 12 group disappeared completely. Histologically, the tumors from 7721/hscIL- 12 without hPBL treatment had numerous lymphocyte infiltration, the tumor cells displayed depression looking, atrophy, focal necrosis and apoptosis , whereas the tumors of 7721 and 7721/pcDNA groups grew thrivingly. Conclusion. hscIL- 12 transduced 7721 cells could induced significant antitumor immune response which resulted in tumor regression totally when the hPBL was inoculated, and also hscIL- 12 has certain effects on mice immune system. These findings suggest that hscIL- 12 and hscIL- 12 gene therapy might have promising prospects in clinical application.

Hepatitis B virus envelope as a targeting gene transfer vector for hepatic cancer cells

Objective： The aim of the study was to observe the transfection efficacy of hepatitis B virus envelope （HBVE） and evaluate its ability as a gene transfer vector for liver cancer cells. Methods： To obtain HBVE, the supematant fluid of HepG 2.2.15 cells was mixed with a PEG8000 solution for concentration and was inactivated by β-propiolactone. The acquired HBVE was used to pack plRES2-EGFP to test its package ability. Then, we examined its quantity and quality with ELISA, PCR, SDS-PAGE and electron microscopy. The plRES2-EGFP was packed with HBVE and obtained the product HBVE-GFP. The plRES2-EGFP was packed with liposome and obtained the product liposome-GFP. HBVE-GFP and liposome-GFP were used to transfer HepG 2 cells to study the transfection efficiency. HBVE-GFP was used to transfer HepG 2, A549, HeLa and FB cells to study the targeting ability. The green fluorescent protein （GFP） expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometry. Results： 1. The acquired HBVE could retain the surface protein HBsAg ＋ pre S1 ＋ pre S2 and had no virus DNA. It had good package ability for plRES2-EGFP. 2. Transfection efficiency： The GFP could be observed in both the liposome group and HBVE group under the fluorescent microscope. But the HBVE group had a higher fluorescent intensity than liposome group. The transfection rate of liposome group was 49.97% ＋ 2.37% while the HBVE group was 70.65% ＋ 3.15% and the fluorescent intensity of the HBVE group was 3-4 times （P = 0.000） for liposome group with the determination of flow cytometry. 3. Targeting ability： The GFP could be observed in the four groups under the fluorescent microscope. The HepG 2 group had the highest fluorescent intensity among the four groups. The transfection rate of HepG 2 group was 71.35% ＋ 0.03% which was highly expressed than other groups （P = 0.000） and the fluorescent intensity of the HepG 2 group was 2-3 times （P = 0.000） for the other 3 groups with the determination of flow cytometry. Conclusion： HBVE can be constructed successfully with the methods of PEG8000 and β-propiolactone from the supernatant fluid of HepG 22.15 cells. The HBVE can be a candidate gene transfer vector for liver cancer cells.

Polymorphism of glutathione S-transferase mu 1 and theta 1 genes and hepatocellular carcinoma in southern Guangxi, China

AIM: Glutathione S-transferase mu 1 (GSTM1) and theta 1 (GSTT1) genes are involved in the metabolism of a wide range of carcinogens, but deletions of the genes are commonly found in the population. The present study was undertaken to evaluate the association between GSTM1 and GSTT1 gene polymorphisms and hepatocellular carcinoma (HCC) risk. METHODS: The genetic polymorphisms were studied at an aflatoxin highly contaminated region in Guangxi, China. Pdymerase chain reaction (PCR) technique was used to detect the presence or absence of the GSTM1 and GSTT1 genes in blood samples. The case group was composed of 181 patients of HCC identified by the pathologists and the control group was composed of 360 adults without any tumor. RESULTS: The frequencies of GSTM1 and GSTT1 null genotypes in the control were 47.8% and 42.7%, while those in the HCC group were 64.6% and 59.7%, respectively. The differences between HCC group and control group were very significant (P<0.01). GSTM1 and GSTT1 combined null genotypes in HCC group and control group were 38.2% and 18.5% respectively, and the difference was significant (P<0.05). CONCLUSION: The GSTM1 and GSTT1 null genotypes are associated with an increased risk of HCC in a special geographic environment. Combination of the two null genotypes in an individual is substantially increased twice the risk of HCC.

Inhibiting effect of antisense hTRT on telomerase activity of human liver cancer cell line SMMC-7721

Objective: To induce changes in biological character of human liver cancer cell line SMMC-7721 by blocking the expression of telomerase genes hTRT and to explore its value in cancer gene therapy. Methods: The vehicle for eukaryotic expression of antisense hTRT was constructed and then transfected into SMMC-7721 cells. The effects of antisense hTRT gene on telomerase activity, cancer cell growth and malignant phenotypes were analyzed. Results: The obtained transfectants that could express antisense hTRT gene stably showed marked decrease in telomerase activity; the shortening of telomere was obvious; cells presented contact growth inhibition; in nude mice transplantation, the rate of tumor induction dramatically decreased. Conclusion: Antisense hTRT gene expression can significantly inhibit telomerase activity of cancer cells and decrease malignant phenotypes in vitro and in vivo. Therefore, as a telomerase inhibitor, antisense hTRT gene may be a new pathway for cancer therapy.

Effect of retinoid X receptor alpha (RXRα) transfection on the proliferation and phenotype of rat hepatic stellate cells in vitro

Objective To study the effect of retinoid X receptor alpha (RXRα) transfection plus treatment with the RXRα ligand, 9-cis-RA, on the proliferation and phenotype of platelet-derived growth factor (PDGF)-activated hepatic stellate cells (HSCs). Methods PDGF activated rat hepatic stellate cells were transfected with eukaryotic expression vector pcDNA3.1- human RXRα, and confirmed by Western blot. Proliferation of transfected HSC was assayed by bromodeoxyuridine (BrdU) incorporation as well as MTT, and the phenotype (α-smooth muscle actin, desmin) was observed by immunocytochemistry with image analysis. Results Transfection of the RXRα gene and treatment with ligand 9-cis-RA of PDGF-activated HSCs extended the increased expression of RXRα protein for at least 168 hours. Cell proliferation and expressions of alpha-smooth muscle actin (α-SMA) and desmin were blocked, compared with groups of sham-transfected, PDGF-activated, no transfection, no ligand treatment, and irrelevant ligand treated HSCs. Conclusion Transfection with the RXRα gene followed by 9-cis-RA ligand treatment will inhibit the proliferation and reverse the phenotype of activated HSC.

Overexpression of bcl-2 protects hepatoma cell line HCC-9204 from ethanol-induced apoptosis

Objective To investigate the effect of overexpression of bcl-2 on ethanol-induced apoptosis of primary hepatocellular carcinoma (HCC) cells.Methods The retrovirus expression vector pDOR-SB containing human bcl-2 cDNA was introduced into a human HCC cell line HCC-9204 by liposome-mediated transfection. pDOR-transfected and non-transfected HCC-9204 cells were used as controls. The expression of Bcl-2 protein by transfected HCC-9204 cells was detected by the immunohistochemical method. Then the cells were cloned with the limited dilution method continually until a monoclonal cell strain whose positive rate of Bcl-2 protein was 100% detected by flow cytometry was obtained. The killing rates of cells were detected by Methabenzthiazuron assay after the treatment of 6% ethanol for 6?h. The extent of apoptosis was analyzed by transferase-mediated dUTP nick end labeling (TUNEL) staining and flow cytometry.Results Most of the pDOR-SB-transfected cells demonstrated Bcl-2 positive signals, while no signal was found in the controls. The positive rate of Bcl-2 protein detected by flow cytometry in the obtained monoclonal cell strain, which was named HCC-bcl2, was 100% after the cells had been cloned 3 times continually. The killing rate, TUNEL index and the scale of sub-G 1 apoptotic peak in HCC-bcl2 cells were all significantly lower than those in the control cells.Conclusion Overexpression of Bcl-2 protein suppresses ethanol-induced apoptosis of the HCC cell line HCC-9204.

Anti-tumor effect of human endostatin mediated by retroviral gene transfer in nude mice

OBJECTIVE: To explore the inhibitory effect of human endostatin gene mediated by retroviral vector on the growth of human liver carcinoma. METHODS: A recombinant retroviral plasmid containing human endostatin gene and signal peptide was engineered and transferred into PA317 cells to produce retrovirus. Human liver carcinoma cells (SMMC7721) were infected with the above retrovirus to build a stable endostatin-transfected liver carcinoma cell line (SMMC-endo). The control liver carcinoma cell line (SMMC-pLncx) was developed in a similar way except that the plasmid was replaced by an empty retroviral vector. Immunohistochemistry and Western blot were used to test the expression and secretion of human endostatin. The biological activity of the expressed human endostatin was assessed by endothelial cell proliferation assay. The growth rates of SMMC-endo and control SMMC-pLncx cells in vivo and in vitro were also observed. RESULTS: The expression and secretion of human endostatin by endostatin-transfected SMMC-endo cells were confirmed by immunohistochemistry and Western blot. Compared with the control group, concentrated supernatant of SMMC-endo cells remarkably inhibited the proliferation of human umbilical vein endothelial cells by 48%, significantly higher than the inhibition by the control (10.2%; P

Metastasis-related genes in hepatocellular carcinoma cell-lines are clustered on chromosome territories predicted by transcriptome and proteome

Recently, we have analyzed expressional transcriptome and proteome for a number of cancer cell lines, including 8 hepatocellular carcinoma （HCC） ones [1,2]. These 8 HCC cell lines consist of 6 metastatic （all with mutant p53） and 2 non-metastatic （with a mutant and a lost p53） ones, being all HBV traceable for relatively close genetic-backgrounds. We have analyzed the related dataset and obtained transcrip- tome （16353 genes） and proteome （7861 proteins） of these 8 HCC cell lines, and explored a group of up-regulated and down-regulated genes [3].