胃癌及癌旁正常组织中微小RNA表达谱的检测

目的 应用微小RNA(miRNA)芯片技术筛选胃癌及癌旁正常组织中差异表达的miRNAs,从中发现与胃癌发生发展相关的miRNAs.方法 收集14例新鲜胃癌组织及癌旁正常黏膜组织,从标本中分离出小RNA,并用Cy3、Cy5双色荧光标记,将标记的小RNA在miRNAs芯片上进行杂交反应.结果 软件MeV4.0对芯片数据进行聚类分析,筛选获得92个在胃癌及癌旁正常组织中差异表达的miRNAs(P＜0.05).相对于癌旁组织,在胃癌组织中有8个miRNAs表达显著上调,11个miRNAs表达显著下调(P＜0.01).结论 胃癌组织和癌旁正常组织中存在差异表达的miRNA分子,它可能与胃癌的发生发展有关.

Abstract：

Objective To screen and identify the microRNA (miRNA) differential expression profiles in gastric cancer and paired adjacent normal tissue by the miRNA microarray technique. Methods Forteen gastric cancer fresh tissue samples and paired adjacent normal mucosa tissue samples were collected. Small RNA was isolated from the samples, labeled with Cy3, Cy5 two-color fluorescence and hybridized on miRNA microarray. Results By analysis of Software MeV4. 0 based on microarrays screening, 92 gastric cancer related miRNAs (P ＜ 0. 05 ) were obtained. In the gastric carcinoma, compared with paired adjacent normal tissue, 8 miRNAs were significantly up-regulated and 11 miRNAs were significantly downregulated (P ＜ 0. 01 ). Conclusion MiRNAs are differentially expressed between gastric carcinoma and adjacent normal tissue, which may be related to the pathogenesis and progression of gastric cancer.     

肝肠钙黏蛋白在胃癌中的表达及其与患者预后的关系

Objective To explore the expressiom of liver-intestine (CDH17) in gastric cancer and its correlation with prognosis. Methods CDH17 protein expression was detected by immunohistochemistry on tissue microarray, which containing 264 samples of primary gastric cancer and the corresponding paracancerous tissue,and 104 paired lymph node metastases (LNM). Results CDH17 was up-regulated in gastric cancer compared with paired nomal gastric mucosa and correlated with clinical stage,invasion depth,LNM,distant metastasis. Immnoreactivity of CDH17 was significantly higher in LNM than in primary gastric cancer and normal mucosa. There was a negative correlation between the high expression of CDH17 and overall survival (OS) or disease free survival (DFS) of gastric cancer patients. CDH17 was an independent prognostic factor for OS [hazard ratio ( HR) 3. 97 ; 95% confidence interval ( CI) 1. 39-10. 28; P ＜ 0. 05] and DFS (HR 2. 32 ;95% CI 1.17-4. 52; P ＜ 0. 05). Conclusion CDH17 may play an important role in gastric cancer progression and could serve as a biomarker to identify patients at risk of metastasis or recurrence after gastrectomy.     

肝肠钙黏蛋白在胃癌中的表达及其与患者预后的关系

Objective To explore the expressiom of liver-intestine (CDH17) in gastric cancer and its correlation with prognosis. Methods CDH17 protein expression was detected by immunohistochemistry on tissue microarray, which containing 264 samples of primary gastric cancer and the corresponding paracancerous tissue,and 104 paired lymph node metastases (LNM). Results CDH17 was up-regulated in gastric cancer compared with paired nomal gastric mucosa and correlated with clinical stage,invasion depth,LNM,distant metastasis. Immnoreactivity of CDH17 was significantly higher in LNM than in primary gastric cancer and normal mucosa. There was a negative correlation between the high expression of CDH17 and overall survival (OS) or disease free survival (DFS) of gastric cancer patients. CDH17 was an independent prognostic factor for OS [hazard ratio ( HR) 3. 97 ; 95% confidence interval ( CI) 1. 39-10. 28; P ＜ 0. 05] and DFS (HR 2. 32 ;95% CI 1.17-4. 52; P ＜ 0. 05). Conclusion CDH17 may play an important role in gastric cancer progression and could serve as a biomarker to identify patients at risk of metastasis or recurrence after gastrectomy.     

Association of hypoxia-inducible factor 1α expression with microlymphatic vessel density and lymph node micrometastasis in rectal cancer

Objective To explore the association of the expression of hypoxia-inducible factor 1α (HIF-1α) with microlymphatic vessel density (MLVD) and lymph node micro-metastasis in rectal cancer.Methods The experimental group consisted of 40 middle-low rectal cancer specimens pathologically confirmed at the First Affiliated Hospital of Anhui Medical University between 2000 and 2003.Forty samples of normal tissues taken from the corresponding area around the cancer were used as the control group. Immunohistochemistry was used to detect HIF-1α expression and MLVD in both the tumor tissues and the adjacent normal tissues. Lymph node micrometastasis was ascertained using immunohistochemical staining with CK20. Results In rectal cancer tissues, the HIF-lα expression was 77 386±14 911 and MLVD was 7.3±0.7, significantly higher than those in normal adjacent tissues(33 092±5877 and 0.3±0.2, both P＜0.01). The HIF-1α expression was positively correlated with MLVD in rectal cancer (r=0.781, P＜0.01). Thirty-one patients had no lymph nodes metastasis and 10 had micrometastasis. The HIF-1α expression and MLVD in specimens with lymph node micrometastasis was significantly higher than that in those without lymph node micrometastasis(P＜0.05). Conclusion HIF-1α and MLVD play important roles in the development of rectal cancer, which may promote lymphatic micrometastasis in rectal cancer.     

PHF1O磷酸化多克隆抗体的制备与其在胃癌细胞PHF10磷酸化水平检测中的应用

目的 制备PHD指蛋白 10(PHF10)磷酸化多克隆抗体并用于PHF10蛋白的研究,探讨其在胃癌发生发展中的作用与机制.方法 结构预测后应用固相法合成磷酸化多肽.应用磷酸化多肽免疫家兔得到多克隆抗体,并利用Dotblot方法进行此抗体特异性效价的鉴定,结果显示PHF10最低阳性检测浓度为7.8μg/L.利用PHF10磷酸化多抗对8株胃癌细胞和胃黏膜永生化细胞PHF10蛋白磷酸化水平进行检测,胃癌细胞株PH F10磷酸化水平明显高于胃黏膜永生化细胞GES-1.结果 制备出的PHF10蛋白磷酸化多克隆抗体效价较高,可用于磷酸化PHF10蛋白的鉴定.结论 此多克隆抗体可用于PHF10蛋白的研究.

Abstract：

Objective To prepare the phosphorylational polyclonal antibody to PHD finger protein 10 (PHF10) and apply it to the study on PHF10 in the tumorigenesis and development of primary gastric cancer.Methods Phosphorylated polypeptide of PHF10 was synthesized by GL Biochem Ltd ( Shanghai).Phosphorylational polyclonal antibody was obtained by immunizing rabbits with the polypeptide.The quality of the antibody was identified by Dot blotting.The minimal detectable concentration of the antibody to PHF10 was 7.8 μg/L.Results The phosphorylational polyclonal antibody to PHF10 was obtained and proved to be effective.By using the phosphorylational polyclonal antibody to PHF10,the phosphorylation status of PHF10 was detected in gastric cancer cells and immortalized gastric epithelial cells,GES-1.The results showed phosphorylation status in PHF10 in gastric cancer cells was higher than that in immortalized gastric epithelial cells,GES-1.Conclusion The phosphorylational polyclonal antibody could be applied to the research on phosphorylation status in PHF10 and phosphorylated PHF10 was expressed higher in gastric cancer cells than in GES-1 but varied in intensity.     

肝肠钙黏蛋白在胃癌中的表达及其与患者预后的关系

目的 探讨肝肠钙黏蛋白(CDH17)在胃癌中的表达及其与预后的关系.方法 应用组织芯片和免疫组织化学方法检测264例胃癌、正常黏膜及104例转移淋巴结中CDH17的表达.结果 CDH17在胃癌中的表达率高于正常黏膜(63.26%比9.85%,P＜0.001),与临床分期(P＜0.01)、浸润深度(P＜0.01)、淋巴结转移(P＜0.01)和远处转移(P＜0.05)相关.转移淋巴结的CDH17表达率(81.73%)高于原发癌和正常黏膜(P＜0.001).CDH17表达与术后总体生存率和无瘤生存率负相关,CDH17是影响总体生存[风险比(HR)3.97;95%可信区间(CI)1.39～10.28;P＜0.05]和无瘤生存(HR 2.32;95%CI 1.17～4.52;P＜0.05)的独立预后因素.结论 CDH17在胃癌进展中起重要作用,可作为预测术后转移复发的标记物.

Abstract：

Objective To explore the expressiom of liver-intestine (CDH17) in gastric cancer and its correlation with prognosis. Methods CDH17 protein expression was detected by immunohistochemistry on tissue microarray, which containing 264 samples of primary gastric cancer and the corresponding paracancerous tissue,and 104 paired lymph node metastases (LNM). Results CDH17 was up-regulated in gastric cancer compared with paired nomal gastric mucosa and correlated with clinical stage,invasion depth,LNM,distant metastasis. Immnoreactivity of CDH17 was significantly higher in LNM than in primary gastric cancer and normal mucosa. There was a negative correlation between the high expression of CDH17 and overall survival (OS) or disease free survival (DFS) of gastric cancer patients. CDH17 was an independent prognostic factor for OS [hazard ratio ( HR) 3. 97 ; 95% confidence interval ( CI) 1. 39-10. 28; P ＜ 0. 05] and DFS (HR 2. 32 ;95% CI 1.17-4. 52; P ＜ 0. 05). Conclusion CDH17 may play an important role in gastric cancer progression and could serve as a biomarker to identify patients at risk of metastasis or recurrence after gastrectomy.     

胰腺癌组织中程序性细胞死亡因子4与细胞色素C的关系

Objective To study the relationship between the expression of cytochrome c ( Cyt c) and programmed cell death 4 (PDCD4) in pancreatic cancer, and investigate the pathway of PDCD4 inducing the apoptosis of pancreatic cancer cells. Methods Pancreatic cancer specimens from 69 patients who received pancreatic resection from 1990 to 2002 in First Affiliated Hospital of China Medical University were collected. The expression of Cyt c in the 69 paraffin specimens of pancreatic cancer was detected by immunohistochemistry, and the expression of Cyt c in 8 samples of cold-preserved fresh pancreatic cancer and normal pancreatic tissues were detected by Western blot. The expression of PDCD4 and Cyt c in pancreatic cancer was analyzed by paired t test and chi-square test. Results Compared with normal pancreatic tissues, the expression of Cyt c in pancreatic cancer was significantly decreased. The positive expression rate of Cyt c in 69 samples of pancreatic cancer was 41% (28/69). The expression of Cyt c was positive in most patients with positive expression of PDCD4, and the expression of PDCD4 was negative in most patients with negative expression of Cyt c. The expression of PDCD4 and Cyt c was closely correlated with each other (χ2= 10.52, P < 0.05). Conclusions There is a close relationship between the expression of PDCD4 and Cyt c in pancreatic cancer. PDCD4 may induce the apoptosis of pancreatic cancer cells through mitochondrial pathway.     

Hedgehog信号通路在脑胶质瘤的表达及其预后意义

目的 观测Hedgehog信号通路在脑胶质瘤的表达,探讨其表达在脑胶质瘤的预后意义.方法 选取118例原发性脑胶质瘤患者的手术切除标本,运用免疫组织化学方法检测Sonic hedgehog(Shh)、受体Patched(Ptch)及下游转录因子Gli1的表达,采用Kaplan-Meier生存分析和Cox比例风险回归模型评价脑胶质瘤患者的预后.结果 免疫染色结果显示Shh、Ptch和Gli1的阳性表达率随胶质瘤病理等级升高呈增强趋势(P＜0.01);随KPS评分的下降而成增强趋势(P＜0.01).生存分析表明,阳性表达Shh、Ptch和Gli1的胶质瘤患者总体存活率低于三者不表达的患者(P＜0.01).多因素Cox分析显示KPS(P＜0.05)、WHO grade(P＜0.01)、Shh(P＜0.05)、Ptch (P＜0.05)和Gli1(P＜0.05)是影响脑胶质瘤预后的独立因素.结论 脑胶质瘤的Shh-Ptch1-Gli1 信号通路处于激活状态,与脑胶质瘤的临床病理特征及预后参数密切相关,提示Hedgehog信号通路的活化在脑胶质瘤的恶性潜能和患者的生存时间起重要的预示作用.     

p53上调促凋亡因子在胃癌组织中的表达及其临床意义

目的 研究p53 上调促凋亡因子(p53 up-regulated modulator of apoptosis,PUMA)在胃癌绀织中的表达及其与临床病理学指标的关系.方法 应用组织芯片和免疫组化Envision方法 检测84例胃癌及配对的癌旁正常胃黏膜组织中PUMA和p53蛋白表达;分析PUMA表达与临床病理学指标的关系;分析PUMA表达与p53表达的相关性.结果 PUMA在胃癌组织中的阳性率(33%)低于癌旁正常胃黏膜组织(62%),两者之间筹异有统计学意义,P<0.01;PUMA的表达与肿瘤浸润深度、pTNM分期及预后有关(P<0.05).p53在胃癌组织中的表达(45.2%)显著高于癌旁正常胃黏膜组织(0),两者之间差异有统计学意义,P<0.01;PUMA与p53表达有显著的相关性(P<0.05).结论 PUMA低表达与胃癌浸润深度、胃癌中、晚期及预后不良相关,有可能成为胃癌预后的预测指标.     

甲状腺乳头状癌组织中Shh和Gli1的表达及其临床意义研究

目的 通过检测Sonic Hedgehog信号通路关键分子Shh和Gil1在甲状腺乳头状癌表达的情况,探讨其与甲状腺乳头状癌临床病理特征的关系及临床意义.方法 应用免疫组织化学方法检测142例甲状腺乳头状癌患者癌组织及其癌旁组织病理蜡块标本中Shh和Gli1的表达情况,分析其与临床病理特征的关系.结果 Shh主要表达于...     

p27Kip1蛋白在胃腺癌中的表达及基与肿瘤细胞增殖的关系

目的 探讨细胞周期素依赖性激酶抑制蛋白27(p27kip1)在胃腺癌中的表达及其对肿瘤细胞增殖的影响和临床意义.方法 采用免疫组织化学SP法检测100例胃腺癌组织中p27kip1蛋白的表达,并与20例正常胃黏膜组织作对照,同时应用流式细胞仪对胃腺癌S期细胞比(SPF)进行检测.结果 正常胃黏膜组织中p27kip1蛋白高表达率明显高于胃腺癌组(X2=15.45,P＜0.01).在高/中分化组,黏膜层/黏膜下层组和无淋巴结转移的病例中,p27kip1蛋白高表达率分别为65.31%、80.00%、73.53%;SPF值分别为21.6±4.2、20.3±3.9、20.1±4.0.但随着胃癌分化程度的降低,侵及肌层/浆膜层或发生淋巴结转移,p27kip1蛋白高表达率明显降低(41.18%、48.24%、42.42%)(X2值分别为5.84、5.16、8.72,P＜0.05或P＜0.01);而SPF值明显升高(25.9±5.1、24:4±4.8、25.7±5.1)(t值分别为4.592、3.127、5.575,P均＜0.01).p27kip1蛋白高表达与SPF值呈显著负相关(t=3.478,P＜0.01).结论 p27kip1与胃腺癌发生发展密切相关.p27kip1蛋白表达降低和高的SPF值常提示胃腺癌的低分化、高侵袭转移能力,提示p27kip1和SPF值可作为临床判断胃腺癌预后的指标.     

Significance of sonic hedgehog signaling after massive hepatectomy in a rat

Purpose

To clarify the functional involvement of hedgehog signaling, especially sonic hedgehog (Shh) and glioma-associated oncogene (Gli)-1 which are known to play an important role in embryonic development and cancer, in the regeneration of a hepatectomized rat liver.

Methods

Six-week-old male Wistar rats were subjected to 70 or 90 % hepatectomy (Hx). Animals were killed at 24, 48 and 72 h after Hx. The liver/body weight ratio was measured as an index of regeneration. Formalin-fixed liver samples were embedded in paraffin, stained for immunohistochemistry with proliferating cell nuclear antigen (PCNA) antibody, and the labeling index was calculated. Immunohistochemistry was also performed with Shh and Gli-1 antibodies.

Results

The liver/body weight ratio gradually increased in both the 70 and 90 % Hx, groups. The hepatocytes were strongly stained for PCNA at 24 h after Hx. Non-parenchymal cells were gradually stained by PCNA from 24 to 72 h after Hx. Shh and Gli-1 expression in hepatocytes was higher after 24 h than at other times and then gradually decreased. Shh and Gli-1 expression in non-parenchymal cells increased gradually, and was found mainly in liver zone I at 72 h after 70 and 90 % Hx.

Conclusions

The expression of both markers suggested that Shh signaling contributes to tissue reconstruction after Hx.     

结直肠癌表观沉默蛋白Bmi1与Notch信号通路调控的关系

目的 探讨人结直肠癌中表观沉默蛋白Bmi1和Notch1的蛋白表达与结直肠癌病理特征的关系,观察Notch通路对结直肠癌细胞增殖凋亡及Bmi1表达的影响。方法 应用免疫组织化学技术(SP法)检测85例结直肠癌组织及其邻近正常肠黏膜中Notch1及Bmi1的蛋白表达;将Notch1通路中γ-分泌酶抑制剂DAPT,作用于结肠癌SW480细胞株,运用噻唑蓝(MTT)比色法检测细胞增殖状态;流式细胞仪观察其对细胞凋亡的影响,Western blot检测ICN及Bmi1蛋白的表达。结果 结直肠癌组织中Notch1和Bmi1蛋白表达的阳性率明显高于正常肠黏膜(P＜0.05),分别为61.2％ (52/85)对15.3％ (13/85)和56.5％ (48/85)对17.6％ (15/85);Bmi1表达率与Notch1表达率呈正相关(r =0.625,P＜0.01),并与肿瘤分化程度、分期及淋巴结转移有关(P＜0.05);阻断Notch1通路(DAPT)可抑制SW480细胞的增殖,诱导其凋亡,作用12、24、36 h后其增殖抑制率分别为13.1％、17.5及22.6％,而凋亡率为32.7％、45.6％及67.2％;同时Notch1胞内活性段ICN随作用时间延长而下降,而Bmi1表达水平也逐渐降低。结论 结直肠癌中Bmi1与Notch1表达密切相关,阻断Notch通路可抑制Bmi1表达,同时可抑制结肠癌细胞的增殖,促进其凋亡。