定量免疫PCR检测血清胃肿瘤相关MGAgs新方法的建立及在胃癌早期诊断中的预警价值

建立定量免疫PCR检测血清胃肿瘤相关MGAgs新方法及研究其在胃癌早期诊断中的预警价值。采用定量免疫PCR技术检测43例胃癌、37例健康献血员血清中MGAgs的含量。研究显示,血清MGAgs含量在健康对照组与胃癌组组间存在显著差异(P<0.01),应用定量免疫PCR方法检测50μL血清MGAgs的截断值相当于为5972个MKN45细胞裂解所得抗原。研究表明,定量免疫PCR技术检测血清MGAgs抗原具有较高的灵敏性和稳定性,通过对胃癌患者血清中MGAgs进行定量检测,可能用于胃癌的诊断和鉴别诊断。

Expression of matrix metalloproteinases 2 and 9 in human gastric cancer and superficial gastritis

AIM:To assess expression of matrix metalloproteinases 2(MMP2)and MMP9 in gastric cancer,superficial gastritis and normal mucosa,and to measure metalloproteinase activity.METHODS:MMP2 and MMP9 mRNA expression was determined by quantitative real-time polymerase chain reaction.Normalization was carried out using three different factors.Proteins were analyzed by quantitative gelatin zymography(qGZ).RESULTS:18S ribosomal RNA(18SRNA)was very highly expressed,while hypoxanthine ribosyltransferase-1(HPRT-1)was moderately expressed.MMP2 was highly expressed,while MMP9 was not detected or lowly expressed in normal tissues,moderately or highly expressed in gastritis and highly expressed in cancer.Relative expression of 18SRNA and HPRT-1 showed no significant differences.Significant differences in MMP2 and MMP9 were found between cancer and normal tissue,but not between gastritis and normal tissue.Absolute quantification of MMP9 echoed this pattern,but differential expression of MMP2 proved conflictive.Analysis by qGZ indicated significant differences between cancer and normal tissue in MMP-2,total MMP-9,250 and 110 kDa bands.CONCLUSION:MMP9 expression is enhanced in gastric cancer compared to normal mucosa;interpretation of differential expression of MMP2 is difficult to establish.

MicroRNA Expression Signature in Gastric Cancer

Objective： To identify the miRNA specific signature as novel diagnostic and prognostic tools for gastric cancer. Methods： miRNAs expression profiling of 3 normal gastric tissues, 24 malignant tissues, gastric cancer cell SGC7901 and normal gastric cell GES-1 were detected using microarray technology. The hierarchical clustering algorithm of the Cluster software was used to analyse the miRNAs expression of all samples. The expression levels of miR-433 and miR-9 which were significantly down-regulated in gastric cancer tissues and SGC7901 cells by microarray technology were validated by quantitative Real-time PCR （qRT-PCR）. Results： Differential expressions of 26 individual miRNAs between normal samples （including 3 normal gastric tissues and GES-1 cells） and carcinomas （including 24 malignant tissues and SGC7901 cells） were discovered, 19 of them showing down-regulation and 7 up-regulation in carcinoma samples. Hierarchical clustering of the Cancer samples by their miRNA expression accurately separated the carcinomas from normal samples and further their histotypes of carcinomas. The expression levels of miR-433 and miR-9 were significantly down-regulated in gastric cancer tissues and SGC7901cells Conclusion： The differential expression of miR-433 and miR-9 may be used as a novel diagnostic tool for gastric cancer.

Detection of carcinoembryonic antigen mRNA in peritoneal washes from gastric cancer patients and its clinical significance

瞄准：建立一个更敏感的方法因为在腹的免费癌症房间的察觉在外科期间从胃的癌症病人洗并且评估它的临床的意义。方法：在腹的 carcinoembryonic 抗原(CEA ) mRNA 层次从胃的癌症的 65 个盒子洗被即时 RT-PCR 检测。腹洗室年龄细胞学(PLC ) 同时被用于免费癌症房间的察觉。包括的腹从 5 个成年健康志愿者从良性的胃的疾病和血样品的 5 个盒子洗的否定控制。结果：在腹没有 CEA mRNA 从良性的胃的疾病病人并且在成年健康志愿者的血洗。即时 RT-PCR 检测的免费癌症房间的积极百分比是 47.7% ，仅仅， 12.3% 显著地被 CEA mRNA 的 PLC.The 积极的率与在腹转移和胃的癌症的舞台之间的浆膜侵略联系。结论：实时 RT-PCR 是一个敏感、快速的方法因为在腹的免费癌症房间的察觉洗。在腹的免费癌症房间的存在洗与胃的癌症的病理学的阶段有关。

Effects of Wei Chang An on expression of multiple genes in human gastric cancer grafted onto nude mice

AIM:To investigate the expression of multiple genes in Chinese jianpi herbal recipe Wei Chang An (WCA) in human gastric cancer cell line SGC-7901. METHODS:A human gastric adenocarcinoma cell line SGC-7901 grafted onto nude mice was used as the animal model. The mice were randomly divided into 3 groups, one control and the two representing experimental conditions. Animals in the two experimental groups received either WCA over a 34-d period or 5-fluorouracil (5-FU) over 6-d period starting at 8th d after grafting. Control animals received saline on an identical schedule. Animals were killed 41 d after being grafted. The expression profiles in paired WCA treated gastric cancer samples and the N.S. control samples were studied by using a cDNA array representing 14 181 cDNA clusters. The alterations in gene expression levels were confirmed by Real-time Quantitative polymerase chain reaction (qPCR). RESULTS:When compared with controls, the average tumor inhibitory rate in WCA group was 44.32% ± 5.67% and 5-FU 47.04% ± 11.33% (P < 0.01, respectively). The average labeling index (LI) for PCNA in WCA group and 5-FU group was significantly decreased compared with the control group. Apoptotic index (AI) was significantly increased to 9.72% ± 4.51% using the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate fluorescence nick end labeling (TUNEL) method in WCA group compared with the controls 2.45% ± 1.37%. 5-FU group was also found to have a significantly increased AI compared with the controls. The expression of cleaved Caspase-3 in WCA group and 5-FU group was significantly increased compared with the control group respectively. There were 45 different expressed sequence tags (ESTs) among the control sample pool and WCA sample pool. There were 24 ESTs up-regulated in WCA samples and 21 ESTs down-regulated. By using qPCR, the expression level of Stat3, rap2 interacting protein x (RIPX), regulator of differentiation 1 (ROD1) and Bcl-2 was lower in WCA group than that in control group respectively. By using SP immunohistochemical method the expression of Phospho-Stat3 (Tyr705) and Bcl-2 in WCA group and 5-FU group was significantly decreased compared with the control group respectively.

Myofibrillogenesis regulator-1 overexpression is associated with poor prognosis of gastric cancer patients

AIM: To investigate the expression of myofibrillogenesis regulator-1 (MR-1) in relation to clinicopathological parameters and postoperative survival in a group of Chinese patients with gastric cancer. METHODS: In our previous study of human wholegenome gene expression profiling, the differentially expressed genes were detected in the gastric cancer and its adjacent noncancerous mucosa. We found that MR-1 was associated with the location and differentiation of tumors. In this study, MR-1 protein expression was determined by immunohistochemistry in specimens of primary cancer and the adjacent noncancerous tissues from gastric cancer patients. A set of real-time quantitative polymerase chain reaction assays based on the Universal ProbeLibrary-a collection of 165 presynthesized, fluorescence-labeled locked nucleic acid hydrolysis probes-was designed specifically to detect the expression of MR-1 mRNA. The correlation was analyzed between the expression of MR-1 and other tumor characteristics which may influence the prognosis of gastric cancer patients. A retrospective cohort study on the prognosis was carried out and clinical data were collected from medical records. RESULTS: MR-1 mRNA and protein could be detected in gastric cancer tissues as well as in matched noncancerous tissues. MR-1 was up-regulated at both mRNA (5.459 ± 0.639 vs 1.233 ± 0.238, P < 0.001) and protein levels (34.2% vs 13.2%, P = 0.003) in gastric cancer tissues. Correlation analysis demonstrated that high expression of MR-1 in gastric cancer was significantly correlated with clinical stage (P = 0.034). Kaplan-Meier analysis showed that the postoperative survival of the MR-1 positive group tended to be poorer than that of the MR-1 negative group, and the difference was statistically significant (P = 0.002). Among all the patients with stageⅠ-Ⅳ carcinoma, the 5-year survival rates of MR-1 positive and negative groups were 50.40% and 12.70%, respectively, with respective median survival times of 64.27 mo (95%CI: 13.41-115.13) and 16.77 mo (95%CI: 8.80-24.74). Univariate and multivariate analyses were performed to compare the impact of MR-1 expression and other clinicopathological parameters on prognosis. In a univariate analysis on all 70 specimens, 6 factors were found to be significantly associated with the overall survival statistically: including MR-1 expression, depth of invasion, distant metastasis, lymph node metastasis, vascular invasion and the tumor node metastasis (TNM) stage based on the 7th edition of the International Union against Cancer TNM classification. To avoid the influence caused by univariate analysis, the expressions of MR-1 as well as other parameters were examined in multivariate Cox analysis. Clinicopathological variables that might affect the prognosis of gastric cancer patients were analyzed by Cox regression analysis, which showed that MR-1 expression and TNM stage were independent predictors of postoperative survival. The best mathematical multivariate Cox regression model consisted of two factors: MR-1 expression and TNM stage. Our results indicated that MR-1 protein could act as an independent marker for patient overall survival [Hazard ratio (HR): 2.215, P = 0.043]. CONCLUSION: MR-1 is an important variable that can be used to evaluate the outcome, prognosis and targeted therapy of gastric cancer patients.

Clinical significance of MET gene amplification in metastatic or locally advanced gastric cancer treated with first-line fluoropyrimidine and platinum combination chemotherapy

Objective: To investigate the clinical significance of MET gene amplification in patients with gastric cancer in the palliative setting.Methods: MET amplification was assessed using fluorescence in situ hybridization(FISH) in 50 patients and quantitative polymerase chain reaction(qPCR) in 326 patients;259 patients treated with first-line fluoropyrimidine and platinum were included for survival analysis.Results: The results of FISH and qPCR indicated that the c-MET/CEP7 ratio was correlated with gene copy number. The optimal cutoff value for the copy number using qPCR to detect MET gene amplification with FISH was 5(κ=0.778, P<0.001). Twenty-one out of 326 patients(6.4%) were identified as MET amplification with a copy number of >5 detected by qPCR. MET-amplified gastric cancer was associated with an Eastern Cooperative Oncology Group(ECOG) performance status(PS) score of ≥2(33.3% vs. 10.5% P=0.007), peritoneal metastasis(76.2% vs. 46.2%, P=0.008), and elevated bilirubin levels(28.6% vs. 7.3%, P=0.006). The median overall survival(OS) and progression-free survival(PFS) were 11.9 and 5.6 months, respectively. MET-amplified gastric cancer was not associated with survival outcomes [hazard ratio(HR)=0.68, 95% confidence interval(95% CI): 0.35-1.32,P=0.254 for PFS;HR=0.68, 95% CI: 0.35-1.32, P=0.251 for OS].Conclusions: qPCR can be used to detect MET gene amplification. MET amplification was not a predictor of poor prognosis in patients with metastatic or unresectable gastric cancer.

SYBR Green I Realtime PCR检测胃癌组织中LIN28基因方法的建立

目的建立SYBRGreen I荧光染料实时定量PCR方法，测定LIN28基因在胃癌组织中的表达水平，并进行实验室方法学评价。方法采用RT—PCR扩增LIN28基因片段，将该片段克隆到质粒载体PMD18-T上，转化入大肠埃希氏菌（DH5a）中，PCR鉴定后，提取重组质粒，以阳性质粒为模板，建立SYBR—GreenI荧光定量PCR标准曲线，以pactin为内参，运用Rotor—Gene6000seriessoftware做相对定量分析，检测30例胃癌患者的癌组织中LIN28的表达水平。结果用重组质粒制作的标准曲线循环阂值与模板浓度有良好的线性关系，内参pactin和LIN28标准曲线相关系数分别为0．9997和0．9991，其最低的检测拷贝数为1，LIN28溶解曲线呈单个特异峰。结论成功地构建了LIN28的原核表达载体。建立的SYBRGreenI荧光染料实时定量PCR法特异度、敏感度高，稳定性好，可用于定量测定胃癌组织中LIN28的表达水平。

Reference genes for quantitative RT-PCR data in gastric tissues and cell lines

AIM:To evaluate the suitability of reference genes in gastric tissue samples and cell lines.METHODS:The suitability of genes ACTB,B2M,GAPDH,RPL29,and 18S rRNA was assessed in21 matched pairs of neoplastic and adjacent nonneoplastic gastric tissues from patients with gastric adenocarcinoma,27 normal gastric tissues from patients without cancer,and 4 cell lines using reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR).The ranking of the best single and combination of reference genes was determined by NormFinder,geNorm,BestKeeper,and DataAssist.In addition,GenEx software was used to determine the optimal number of reference genes.To validate the results,the mRNA expression of a target gene,DNMT1,was quantified using the different reference gene combinations suggested by the various software packages for normalization.RESULTS:ACTB was the best reference gene for all gastric tissues,cell lines and all gastric tissues plus cell lines.GAPDH+B2M or ACTB+B2M was the best combination of reference genes for all the gastric tissues.On the other hand,ACTB+B2M was the best combination for all the cell lines tested and was also the best combination for analyses involving all the gastric tissues plus cell lines.According to the GenEx software,2 or 3 genes were the optimal number of references genes for all the gastric tissues.The relative quantification of DNMT1 showed similar patterns when normalized by each combination of reference genes.The level of expression of DNMT1 in neoplastic,adjacent non-neoplastic and normal gastric tissues did not differ when these samples were normalized using GAPDH+B2M(P=0.32),ACTB+B2M(P=0.61),or GAPDH+B2M+ACTB(P=0.44).CONCLUSION:GAPDH+B2M or ACTB+B2M is the best combination of reference gene for all the gastric tissues,and ACTB+B2M is the best combination for the cell lines tested.

血清外泌体MT1-MMP mRNA在胃癌患者中的表达及其与预后的关系

目的 探讨血清外泌体膜1型基质金属蛋白酶(MT1-MMP)mRNA在胃癌患者中的表达及其与预后的关系。方法 选取2016年3月—2018年7月我院行胃癌切除术的98例患者为研究对象。采用实时荧光定量PCR(RT-PCR)测定患者癌组织、癌旁正常组织中MT1-MMP mRNA表达量,分析患者临床病理参数与胃癌组织中MT1-MMP mRNA表达量的关系;对98例患者进行随访,统计术后3年内存活率,比较存活亚组(n=62)与死亡亚组(n=36)患者入组时的胃癌组织与癌旁组织中MT1-MMP mRNA表达水平,运用受试者工作曲线(ROC)分析MT1-MMP mRNA表达水平对术后3年内生存状态的预测效能。结果 98例患者胃癌组织中MT1-MMP mRNA表达水平(0.96±0.12)显著高于癌旁组织(0.80±0.09)(P<0.05);患者胃癌组织中MT1-MMP mRNA表达量与淋巴结转移、浸润程度、TNM分期有关(P<0.05),与性别、年龄、肿瘤位置、肿瘤直径大小及分化程度无关(P>0.05);98例患者术后3年存活亚组入组时的胃癌组织及癌旁组织中MT1-MMP mRNA表达水平显著高于死亡亚组(P<0.05);ROC曲线分析显示,胃癌组织、癌旁组织MT1-MMP mRNA表达水平对患者术后3年生存状态的预测效能有一定局限性(AUC=0.760、0.691),胃癌组织与癌旁组织联合检验的预测效能明显高于单一检测(AUC=0.813)。结论 胃癌组织中MT1-MMP mRNA表达与肿瘤的淋巴转移、TNM分期及浸润程度相关,可一定程度上指导预后。

干细胞标志物Nanog的检测在胃癌诊断中的意义

目的检测胃癌患者癌组织和癌旁组织中干细胞标志物Nanog的表达水平并探讨其与临床病理参数的相关性。方法用RT-PCR和Real-timePCR检测62例胃癌患者癌组织和癌旁组织中Nanog的表达。结果RT-PCR结果显示胃癌组织和癌旁组织中Nanog的表达阳性率分别为58.1%(36/62)、9.7%(6/62),差异有统计学意义(P<0.0001);Real-timePCR结果表明胃癌组织中Nanog相对表达水平高于癌旁组织(P<0.05);胃癌组织中Nanog阳性表达与肿瘤分化状态相关,低、未分化组高于中、高分化组(P<0.05),但与年龄、性别、肿瘤大小、浸润深度、TNM分期和有无淋巴结转移无关。结论Nanog表达与胃癌的发生及其分化状态相关,可作为胃癌诊断的一个新的分子标志。

胃癌组织miRNA差异表达的初步分析

目的分析胃癌组织特异性微小RNA(miRNA)表达谱,为深入研究miRNA与胃癌发生和发展的关系以及寻找新的胃癌分子标志物奠定基础。方法利用miRNA芯片就21对胃癌和癌旁配对正常组织的miRNA表达谱进行检测和初步生物信息学分析,Real-TimePCR验证miRNA芯片检测结果。结果 miRNA芯片检测发现,在胃癌及其癌旁配对正常组织中共有88个差异表达miRNA,其中上调最明显的是miR-21、miR-196b、miR-301a、miR-431*、miR-550*、miR-18a和miR-135b;下调最明显的是miR-139-3p、miR-628-3p、miR-596、miR-99b*和miR-638。Real-TimePCR对其中2个上调(miR-181、miR-19b)和2个下调(miR-134、miR-31)miRNA的验证结果与芯片检测所示具有较好的一致性。结论胃癌组织具有特异性的miRNA表达谱,这些差异表达的miRNA有可能成为新的胃癌分子标志物。

胃癌腹膜转移中GALECTIN-3MRNA的表达与意义

目的探讨GALECTIN-3基因在胃癌腹膜转移中的作用。方法通过实时定量RT-PCR方法,对35例同期伴腹膜转移病人的胃癌原发灶、淋巴结转移灶、腹膜转移灶、正常胃粘膜组织中GALECTIN-3MRNA的表达水平进行实时定量RT-PCR检测。结果胃癌原发灶(385.639±98.534)、腹膜转移灶(368.589±93.431)、淋巴结转移灶(375.920±94.346)中GALECTIN-3MRNA表达量同正常胃粘膜(2.158±0.896)中的表达量相比,差异具有统计学意义(P<0.05),胃癌原发灶、腹膜转移灶及淋巴结转移灶三者中的表达量两两相比,差异无统计学意义(P>0.05)。GALECTIN-3MRNA在胃癌原发灶与腹膜转移灶中的表达呈正相关(R=0.997,P=0.000)。结论GALECTIN-3MRNA在胃癌中的高表达与胃癌腹膜转移密切相关。胃癌病灶的GALECTIN-3MRNA检测可作为术前判断胃癌已伴腹膜转移或具有腹膜转移潜能的生物学标记。

胃癌患者的血清乳酸脱氢酶水平及其基因在癌组织的表达与患者预后的相关性研究

目的：探讨胃癌患者的血清乳酸脱氢酶（lactate dehydrogenase，LDH）水平及癌组织中该基因的表达与患者预后的相关性。方法2012-2013年经某院病理确诊的胃癌患者的术前血清标本及癌组织标本各60例，检测血清标本的LDH水平，并使用实时荧光定量反转录PCR的方法检测60例患者的正常胃组织、癌组织及淋巴结组织中 LDH基因的表达。最后，将血清 LDH水平和胃癌组织中 LDH基因的差异表达与患者的临床病理特征进行相关性分析，其中多样本比较采用单因素方差分析，两样本比较使用独立t检验。结果胃癌患者血清LDH为340.89±10.67 IU/ml，显著高于对照组（t＝24.7，P＜0.01）；胃癌组织及淋巴结组织中 LDH的相对表达量分别是正常组织的3.39及2.35倍。与临床病理特征的相关性分析提示血清 LDH的升高与 pTNM分期及是否有淋巴结转移相关（t＝5.2，4.8，P 值均＜0.01），胃癌组织LDH的高表达与肿瘤大小、pTNM分期及是否有淋巴结转移相关（t＝18.2，15.3，F＝7.2，P值均＜0.01）。结论胃癌患者的血清 LDH水平及其组织中 LDH基因表达显著升高，其高表达与患者预后相关。检测血清 LDH 水平及胃癌组织中LDH表达可能成为评估胃癌患者预后的潜在指标。

长链非编码RNA MEG3在胃癌中的表达及其与预后的关系

目的:研究长链非编码RNA(long non-coding RNA,lnc RNA)母系表达基因3(maternally expressed gene 3,MEG3)在胃癌组织中的表达及其与预后的关系,并进一步探讨MEG3与凋亡相关蛋白P53、鼠双微基因2(murine double minute 2,MDM2)的相关性。方法:收集2012年9月至2013年6月青岛大学附属青岛市市立医院55例行手术治疗的胃癌切除标本(包括癌组织及对应的远端正常组织),实时荧光定量PCR(q RT-PCR)检测胃癌组织中MEG3的表达,并分析其与临床病理参数的关系;应用免疫蛋白印迹法(Western blot)检测胃癌中P53、MDM2蛋白的表达水平,并分析其与MEG3的相关性。结果:lnc RNA MEG3在胃癌组织的相对表达水平(7.98±0.19)低于所对应的正常组织(9.47±0.18,P<0.05);在胃癌组织(r=-0.627,P<0.001)及远端正常组织(r=-0.807,P<0.001)中,P53与MDM2的表达呈负相关;P53与MEG3的表达呈正相关(r=0.591,P<0.05),MDM2与MEG3的表达呈负相关(r=-0.346,P<0.05);MEG3高表达者较低表达者中位生存期明显延长。结论:MEG3在胃癌发生发展过程中起抑癌基因的作用;MEG3与P53、MDM2在胃癌发生发展的生物学机制上有重要联系;检测MEG3的表达水平对胃癌预后有潜在价值。

BMP7基因检测在结直肠癌和胃癌诊断中的意义

目的建立检测骨形态发生蛋白7(BMP7)基因的实时荧光定量RT-PCR法,探讨BMP7对结直肠癌、胃癌的诊断意义。方法构建BMP7基因的标准质粒,建立RT-PCR,real-time RT-PCR等方法检测并比较BMP7基因在21例胃癌组织、19例结直肠癌组织与对应癌旁组织中的表达水平,用免疫组织化学验证BMP7的蛋白水平差异。结果建立的BMP7检测法稳定和准确;19例结直肠癌中16例(84.2%)癌组织表达的BMP7基因显著高于对应的癌旁组织(P<0.05)。21例胃癌组织中11例(52.4%)癌组织表达高于对应癌旁组织,但无显著差异(P>0.05)。免疫组织化学在蛋白水平上证实了肿瘤组织和癌旁组织中BMP7的表达水平差异。结论成功建立了检测BMP7的实时荧光定量PCR方法,BMP7基因可作为结直肠癌检测的新靶点。

应用蛋白质组学技术筛选胃癌耐药相关蛋白质

胃癌多药耐药性是临床胃癌化疗失败最主要的原因之一,但其分子机制仍然不太清楚.为了寻找新的胃癌耐药相关的蛋白质,揭示胃癌多药耐药的分子机制,以胃癌细胞SGC7901和长春新碱诱导的耐药胃癌细胞SGC7901/VCR为研究对象,应用二维凝胶电泳(two-dimensionalelectrophoresis,2-DE)技术分离两种细胞的总蛋白质,图像分析识别差异表达的蛋白质点,基质辅助激光解吸电离飞行时间质谱(matrix-assistedlaserdesorption/ionizationtimeofflightmassspectrometry,MALDI-TOF-MS)及电喷雾电离串联质谱(electrosprayionizationtandemmassspectrometry,ESI-Q-TOF)对差异表达的蛋白质点进行鉴定,蛋白质印迹和实时RT-PCR验证部分差异蛋白质在两株细胞中的表达水平,反义核酸转染技术分析HSP27(heatshockprotein27,HSP27)高表达与SGC7901/VCR耐药的相关性.得到了分辨率较高、重复性较好的两株细胞系的二维凝胶电泳图谱,质谱分析共鉴定了24个差异蛋白质点,蛋白质印迹和实时RT-PCR验证了部分差异蛋白的表达水平,反义寡核苷酸抑制HSP27表达能增加SGC7901/VCR对长春新碱的敏感性.研究结果不仅提示这些差异蛋白质如HSP27,Sorcin等可能与胃癌的多药耐药相关,而且为揭示胃癌细胞的多药耐药性产生机制提供了线索.

隐丹参酮对人胃癌SGC-7901细胞增殖及血管内皮生长因子mRNA表达的影响

目的通过测定隐丹参酮对人胃癌SGC-7901细胞增殖及血管内皮生长因子（VEGF）mRNA的影响,探讨其抗肿瘤的作用机制。方法采用四甲基偶氮唑蓝（MTT）比色法检测隐丹参酮对人胃癌SGC-7901细胞增殖的影响,采用Real Time RT-PCR检测隐丹参酮对VEGF m RNA表达的影响。结果 20、40、60、80、100μmol/L的隐丹参酮作用于人胃癌SGC-7901细胞6-48 h,其生长和增殖均受到一定程度的抑制;24 h为隐丹参酮对SGC-7901细胞抑制作用的最佳时间,IC50为59.11μmol/L;选取40、60和80μmol/L 3个剂量作用于人胃癌细胞SGC7901 24 h后,60和80μmol/L的隐丹参酮均可下调VEGF m RNA的表达（均P〈0.01）。结论隐丹参酮可抑制人胃癌SGC-7901细胞增殖,并能抑制VEGF mRNA的表达,提示这可能是其抗肿瘤的作用机制之一。

黄连素抑制胃癌细胞分子机制的初步探讨

目的初步探讨黄连素(Berberine)抑制胃癌细胞的分子机制。方法利用Real-time PCR检测黄连素对胃癌细胞BGC-823和SGC-7901增殖、凋亡、坏死、自噬及细胞连接相关基因表达水平的影响。结果黄连素处理胃癌细胞后,增殖相关基因IGFBP5、E2F6及MDM2表达水平降低,Notch2和JAG2表达水平升高,而PAK3表达水平无明显变化;凋亡相关基因Bcl-2表达水平降低,Caspase-1及Caspase-8表达水平升高;坏死相关基因RIPK1与RIPK3表达水平降低,自噬相关基因ATG7表达水平升高;细胞连接相关基因Ep CAM表达水平明显降低,E-Cadherin表达水平无明显变化。结论黄连素通过调控增殖、凋亡等相关基因的表达水平发挥抑制胃癌细胞的作用。

Notch1在GES-1和其他胃癌细胞中表达差异性的比较

目的探讨Notch 1在GES-1、AGS、SGC-7901和BGC-823中的表达差异性。方法采用real time RT-PCR技术和Western blot方法研究Notch 1在GES-1、AGS、SGC-7901和BGC-823细胞mRNA和蛋白质水平上的表达。结果与GES-1相比较,Notch 1 mRNA和NOTCH 1蛋白在AGS,SGC-7901和BGC-823中表达显著降低,差异均具有显著性(P<0.01)。结论Notch 1在胃癌的发生中可能作为抑癌基因起作用。