Six1和Six4在食管鳞状细胞癌组织中的表达及其与预后的关系北大核心CSCD

目的探讨Six1和Six4在食管鳞状细胞癌（简称鳞癌）组织中的表达及其与临床病理特征及预后的关系。方法应用组织芯片技术、免疫组织化学EnVision法检测Six1和Six4在292例食管鳞癌组织及其相应的癌旁正常食管上皮组织中的表达水平,分析两者表达与临床病理特征及预后之间的关系。结果在292例食管鳞癌患者中,Six1、Six4蛋白在癌组织的阳性率分别为72．9％（213／292）和56．2％（164／292）,明显高于癌旁正常组织的阳性率,分别为33．2％（97／292）和32．5％（95／292）,差异具有统计学意义（P〈0．05）。X2检验结果显示Six1蛋白表达与肿瘤大小、肿瘤浸润深度及生存状态相关,其在死亡组患者的阳性率82．3％（130／158）,明显高于生存组阳性率61．9％（83／134,P〈0．05）。Six4蛋白表达与肿瘤分化程度及肿瘤浸润深度相关,肿瘤分化程度越高、浸润深度越深其阳性率越高。单因素Log-rank分析显示性别、TNM分期、Six1蛋白表达与食管鳞癌患者的总生存率相关（P〈0．05）。Six1蛋白表达阴性组5年生存率51．9％（41／79）高于阳性组的43．7％（93／213）,差异具有统计学意义（X^2=4．079,P=0．043）,其他临床参数与预后均未显示统计学意义（P〉0．05）。多因素Cox比例风险模型分析显示,TNM分期、Six1蛋白表达水平是影响食管癌术后患者预后的独立因素（P〈0．05）。结论Six1和Six4蛋白在食管鳞癌组织中均高表达,其表达水平与食管鳞癌发生发展及预后密切相关,Six1蛋白高表达是食管鳞癌患者预后不良的因素,可作为临床预测食管鳞癌患者预后的一个重要指标。     

婆罗双树样基因4在人前列腺癌细胞和组织中的表达及其与临床预后之间的关系

 目的：评估婆罗双树样基因4（SALL4）在人前列腺癌细胞系和前列腺癌组织中的表达情况,并明确其表达水平和临床病理学参数间的关系。方法：用免疫荧光技术、RT-PCR和Western blotting检测SALL4在LNCaP、DU145、PC-3细胞系和RWPE-1正常前列腺上皮细胞系中的表达。同时用免疫组化方法检测SALL4在前列腺增生及前列腺癌组织中的表达,并探讨其与Gleason评分等临床病理参数的关系。结果： SALL4蛋白在细胞中主要表达于胞浆,在3种前列腺癌细胞株中SALL4蛋白表达水平要明显高于正常前列腺上皮细胞RWPE-1（P＜005）,而SALL4 mRNA表达水平在4种细胞系中无明显差异（P＞0.05）。免疫组化结果显示SALL4在前列腺癌组织中的表达水平明显高于增生和正常前列腺组织（P＜0.01）。SALL4蛋白表达水平与Gleason评分、前列腺癌临床分期、预后及组织前列腺特异抗原（PSA）表达密切相关,而与患者年龄、治疗前血清总PSA水平、前列腺体积及组织雄激素受体表达无明显相关性。结论：SALL4蛋白在前列腺癌中高表达,提示其在前列腺癌的发生、发展过程中可能起重要作用,有可能成为诊断前列腺癌新的肿瘤标志物及评估其恶性程度、进展和预后的参考指标。     

核转录因子FOXP3蛋白在人脑星形细胞肿瘤组织中的表达及临床意义

目的:检测星形细胞肿瘤组织中FOXP3蛋白的表达,探讨星形细胞肿瘤组织中FOXP3的表达与临床病理特征以及预后的关系。方法:应用免疫组织化学技术检测121例星形细胞肿瘤和5例脑膜瘤,5例正常脑组织中FOXP3的表达。分析FOXP3的表达与星形细胞肿瘤各临床病理参数及预后的关系。结果:FOXP3蛋白在人脑星形细胞肿瘤组织中有表达,在脑膜瘤和正常脑组织中均未见表达,FOXP3蛋白主要表达于间质淋巴细胞的细胞核。在星形细胞肿瘤中,FOXP3的表达在各级别星形细胞肿瘤中表达差异显著(P0.05)。Kaplan-Meier生存分析提示FOXP3阳性组和阴性组生存时间差异显著(P0.01),但COX多因素变量分析结果显示,FOXP3不是影响星形细胞肿瘤患者术后生存时间的独立预后因素(P0.05)。结论:FOXP3在星形细胞肿瘤间质淋巴细胞中的表达与星形细胞肿瘤的恶性程度以及病人的年龄相关。FOXP3阳性的星形细胞肿瘤的预后较FOXP3阴性的星形细胞肿瘤差,但不能作为影响星形细胞肿瘤生存时间的独立预后因素。FOXP3可能成为星形细胞肿瘤免疫调节的新靶点。     

胃癌组织中PD-L1蛋白表达和基因扩增的相关性北大核心CSCD

目的探讨胃癌中程序性死亡分子配体1（PD-L1）蛋白表达和基因扩增的相关性及与临床病理特征的相关关系。方法选取山西省肿瘤医院2011年有完整随访资料和临床病理资料的胃癌患者247例,应用免疫组织化学（IHC）标记及荧光原位杂交（FISH）技术检测胃癌组织中PD-L1蛋白表达和基因扩增情况。结果（1）PD-L1蛋白表达与胃癌临床病理特征的关系：在胃癌组织,肿瘤细胞中PD-L1蛋白表达阳性率为25.9%（64/247）,肿瘤浸润免疫细胞（IC）中PD-L1蛋白表达阳性率为26.7%（66/247）,两者表达具有相关性（P〈0.01）,肿瘤细胞中PD-L1表达与分化程度和肿瘤直径有相关性（P〈0.05）,免疫细胞中PD-L1表达与有无脉管癌栓有相关性（P〈0.05）。（2）PD-L1基因扩增与胃癌临床病理特征的关系：FISH检测PD-L1基因扩增率为19.0%（47/247）。PD-L1基因扩增与年龄、胃大/小弯、肿瘤部位、肿瘤直径及淋巴结转移有相关性（P〈0.05）。（3）IHC检测的肿瘤细胞中PD-L1蛋白表达与FISH检测的PD-L1基因扩增,两种检测方法阳性符合率为25.0%（16/64）,阴性符合率为83.0%（152/183）,总符合率为68.0%（168/247）,IHC和FISH检测结果一致性较差（P＝0.157）。（4）单因素生存分析：肿瘤细胞中PD-L1蛋白表达与预后呈负相关关系,免疫细胞中PD-L1蛋白表达和PD-L1基因扩增与预后均没有明显相关性。脉管癌栓、肿瘤直径、浸润深度和淋巴结转移均是胃癌的不良预后因素（P〈0.05）。（5）多因素Cox回归分析：肿瘤细胞中PD-L1蛋白表达、浸润深度和淋巴结转移均为影响胃癌预后的独立危险因素（P〈0.05）。结论胃癌中PD-L1蛋白表达与基因扩增一致性较差;PD-L1蛋白表达提示预后较差;PD-L1基因扩增与预后无明显相关性。     

PTK7在食管鳞状细胞癌中的表达及临床意义

目的研究酪氨酸蛋白激酶-7（PTK7）在食管鳞状细胞癌、癌旁组织中的表达及其与食管鳞状细胞癌的发展、侵袭、淋巴结转移的关系,分析其在食管鳞癌中的诊断价值及预后变化。方法收集食管鳞状细胞癌标本80例及癌旁组织63例,采用免疫组织化学链霉菌抗生物素蛋白一过氧化物酶连结（SP）法检测PTK7蛋白的表达,结合临床相关因素进行x2检验及Kaplan—meier等统计学分析。结果80例患者食管鳞状细胞癌组织中有45例PTK7蛋白表达阳性（X2=50．17,P〈0．01）,食管癌旁非典型增生及正常鳞状上皮中PTK7蛋白表达均阴性。PTK7表达与食管鳞癌的年龄、性别、分化程度、浸润深度、淋巴结转移、浸润长度、临床分期等比较差异无统计学意义（P〉0．05）;与5年生存率比较差异亦无统计学意义（X2=0．2,P〉0．1）;当肿瘤组织PTK7蛋白表达综合免疫组化评分≥5分时,5年生存率有下降趋势（X2=3．35,P=0．06）。结论PTK7表达与食管鳞癌有关,当PTK7综合免疫评分≥5分时,5年生存率有下降趋势。利用检测PTK7的表达,有助于综合判断食管鳞癌的相关性和临床预后。     

CIC蛋白失表达在少突胶质细胞肿瘤诊断及预后判断中的意义北大核心CSCD

目的探讨CIC蛋白失表达在少突胶质细胞肿瘤中作为预测染色体lp／19q共缺失初筛检测方法的可行性及其对预后的影响。方法回顾性分析首都医科大学宣武医院病理科113例形态学具有少突胶质细胞瘤特征的胶质瘤,分别进行染色体lp／19q共缺失荧光原位杂交（FISH）检测及CIC蛋白免疫组织化学标记。分析CIC蛋白失表达与染色体1p／19q共缺失的相关性,并探讨其与预后的关系。结果在113例组织学诊断为少突胶质细胞肿瘤中,CIC蛋白失表达率为59．3％（67／113）,不同的组织学类型中失表达率不同,在只含有少突胶质细胞成分的肿瘤中失表达率（85．7％．42／49）比混合性的胶质瘤中（39．1％,25／64）高（P〈0．01）。CIC蛋白失表达预测1p／19q共缺失的敏感性和特异性分别为76．1％（54／71）和71．1％（27／38）,假阳性率和假阴性率分别为16．9％（11／65）和38．6％（17／44）。按照2016版（（WHO中枢神经系统肿瘤分类第4版修订版》,本组病例中63例整合诊断符合少突胶质细胞瘤／间变型少突胶质细胞瘤,IDH突变及1p／19q共缺失;CIC蛋白失表达率为81．0％（51／63）,其预测1p／19q共缺失的敏感性和特异性分别提高至81．0％（51／63）和76．9％（20／26）,假阳性率和假阴性率分别降至10．5％（6／57）和37．5％（12／32）。此时,应用Kaplan—Meier法分析CIC蛋白失表达对预后的影响,发现CIC蛋白失表达组比阳性表达组预后稍好,但二者差异无统计学意义（总生存期：P=0．218;无进展生存期：P=0．249）。结论利用免疫组织化学检测CIC蛋白失表达情况,可以作为染色体1p／19q共缺失的一种初筛检测方法。CIC蛋白失表达与预后无关。     

琥珀酸脱氢酶缺陷型胃肠道间质瘤中琥珀酸脱氢酶各亚单位蛋白的表达情况北大核心CSCD

目的探讨琥珀酸脱氢酶（SDH）缺陷型胃肠道间质瘤（GIST）中琥珀酸脱氢酶复合体各亚单位蛋白的表达情况。方法收集贵州医科大学附属医院及四川大学华西医院病理科2003年1月至2017年1月期间经外科切除并经病理学检查证实的GIST标本352例．应用免疫组织化学EnVision二步法检测352例GIST肿瘤组织中琥珀酸脱氢酶各亚单位蛋白的表达情况,对阴性病例进行临床病理特征分析及随访,同时对阴性病例的CKIT基因第9、11、13及17号外显子及血小板源性生长因子受体仪（PDGFRA）基因第12和18号外显子进行扩增并检测。结果352例GIST中SDHB蛋白表达缺陷（阴性）15例,占4．3％（15／352）,其中男性6例,女性9例;首诊年龄15～84岁（中位年龄53岁,平均年龄47岁）;肿瘤发生于胃14例、肠系膜1例;肿瘤直径0．5～15．0cm（平均6．9cm）;上皮样细胞型和混合细胞型各6例,梭形细胞型3例：其中8例在胃壁内呈多结节状生长。4例发生淋巴结转移,1例发生腹腔种植转移,1例发生肝脏、胰腺及淋巴结转移,1例发生脉管内浸润。在SDHB缺陷型GIST中,发现SDHA蛋白表达缺陷（阴性）2例（0．6％,2／352）;未发现SDHC及SDHD蛋白表达缺陷（阴性）病例。共5例获得随访,随访时间为7～151个月（平均56个月）,其中1例于术后8个月死亡（死因不明）;4例无肿瘤转移和复发,患者本人及家族中均未发现副神经节瘤及肺软骨瘤等病史。15例SDH缺陷型GIST均未检测出CKIT及PDGFRA基因突变。结论SDH缺陷型GIST有独特的临床病理特征且预后较好,其中少数为SDHA缺陷。     

Reg3α基因影响胰岛内皮细胞分泌胰岛素

 目的 构建小鼠胰腺损伤模型,利用全基因组芯片技术筛选出高表达基因胰岛再生衍生因子3α（regenerating islet-derived 3 alpha,Reg3α）,然后通过胰岛内皮细胞（MS1）过表达Reg3α,探讨小鼠Reg3α对胰腺再生中的重要作用。 方法 建立小鼠部分（90%）胰腺切除模型,分别于术前、术后12hr、24hr检测血糖, 术后48hr收集小鼠剩余胰腺组织,经全基因组芯片分析、q-PCR验证Reg3α高表达。构建Reg3α过表达质粒,转染MS1细胞系,48hr后ELISA检测细胞上清培养液胰岛素分泌量的变化,并通过q-PCR检测转染后MS1中Pdx1、Insulin2 mRNA表达水平。 结果 比较未转染组(untransfected group)与转染空载体组（PcDNA3.1-transfected group）,MS1上清培养液胰岛素分泌量没有明显变化（14.63±6.88,P＞0.05）;比较转染Reg3α过表达质粒组与未转染组,上清培养液胰岛素分泌量明显升高（193.43±7.09, P＜0.01）;比较转染Reg3α过表达质粒组与转染空载体组,胰岛素分泌量有明显增加（208.07±5.52, P＜0.01）。与未转染组、转染空载体组比较,转染Reg3α过表达质粒后MS1中的Pdx1、Insulin2 mRNA表达水平明显升高（P＜0.01） 结论 胰岛内皮细胞中过表达Reg3α可使Pdx1、Insulin2 mRNA表达水平升高,增加胰岛素的分泌。Reg3α促进内皮细胞表达和分泌胰岛素可能具有对胰岛β细胞功能代偿作用。     

SLP-2在胃癌组织中的表达及其预后意义

 目的：探讨红细胞膜整合蛋白SLP-2（stomatin-like protein 2）在胃癌中的表达及其与临床病理学指标及预后的关系。方法：选取中山大学肿瘤防治中心病理科有完整临床资料的胃癌手术标本190例,应用免疫组织化学方法检测胃癌中SLP-2蛋白的表达,并分析其与临床病理特征及预后的关系。结果：（1）胃癌组织中SLP-2蛋白表达的阳性率为63.2%（120/190）。SLP-2表达与胃癌浸润深度、TNM分期及淋巴结转移有关（P<0.05）,而与患者性别、年龄、肿瘤分化程度、肿瘤直径和远处转移均无关（P>0.05）;（2）Kaplan-Meier 生存曲线分析结果显示：SLP-2阳性表达患者的累积生存率明显低于阴性表达患者（P<0.01）;（3）单因素分析结果显示：SLP-2 的表达、淋巴结转移、肿瘤分化程度、肿瘤直径、TNM分期、浸润深度及远处转移均影响胃癌预后;多因素分析表明,只有肿瘤直径和TNM分期是独立的预后指标。结论：（1）SLP-2在胃腺癌组织中高表达,可能参与胃腺癌的发生发展和转移;（2）SLP-2在一定程度上影响胃癌的预后,其过度表达提示胃癌预后差。     

胃肠道平滑肌肿瘤ras癌基因产物P21表达及其与临床病理的关系

胃肠道平滑肌肿瘤ｒａｓ癌基因产物Ｐ２１表达及其与临床病理的关系蔡建辉，蒋贻康，张玉印，张祥宏，吕贵华我们应用流式细胞术对胃肠道平滑肌肿瘤（ＧＩＳＭＴ）的ｒａｓＰ２１表达进行定量分析。旨在结合组织学观察，探讨判断肿瘤良、恶性及病人预后的新途径。一、材料...     

High expression of RELP (Reg IV) in neoplastic goblet cells of appendiceal mucinous cystadenoma and pseudomyxoma peritonei

The regenerating protein (Reg)-like protein (RELP, also known as Reg IV) is a recently characterized fourth member of the human Reg protein family. The Reg proteins are small, about 20-kD-sized, secretory proteins of C-lectin type. The previously known Reg proteins have been functionally implicated in regeneration, proliferation, and differentiation of the pancreas, liver, and gastrointestinal mucosa. To study the tissue expression of RELP, we raised a monoclonal antibody to RELP. By immunohistochemistry and in situ hybridization, we found a robust de novo expression of RELP in the neoplastic goblet cells of appendiceal mucinous cystadenomas and in the epithelial implants of pseudomyxoma peritonei (PMP). Our findings indicate that RELP serves as a marker for appendiceal mucinous cystadenomas and PMP, and that RELP may contribute to the pathogenesis of these disorders.     

Expression and localization of Reg IV in human neoplastic and non-neoplastic tissues: Reg IV expression is associated with intestinal and neuroendocrine differentiation in gastric adenocarcinoma

Regenerating islet-derived family, member 4 (Reg IV) is a candidate marker for cancer and inflammatory bowel disease. In the present study, immunohistochemical analysis of Reg IV was performed in various human neoplastic (n = 289) and non-neoplastic tissues. In the stomach, foveolar epithelium was negative for Reg IV, whereas goblet cells of intestinal metaplasia and neuroendocrine cells at the base of intestinal metaplasia expressed Reg IV. Neuroendocrine cells of the small intestine and colon showed strong expression of Reg IV, whereas goblet cells of the small intestine and colon showed weak or no expression of Reg IV. Insulin-producing beta cells of the endocrine pancreas were positive for Reg IV. Among 143 gastric adenocarcinomas, Reg IV expression was detected in 42 (29.4%) and was associated with both the intestinal mucin phenotype and neuroendocrine differentiation. No association was found between Reg IV expression and clinical characteristics such as tumour stage and patient prognosis. Of 36 colorectal adenocarcinomas, 13 (36.1%) were positive for Reg IV, which was associated with tumour stage (p = 0.0379, Fisher's exact test). Expression of Reg IV was detected in 14 (93.3%) of 15 colorectal carcinoid tumours. Reg IV expression was also detected in 5 (21.7%) of 23 ductal adenocarcinomas of the pancreas. In contrast, lung cancers (n = 30) and breast cancers (n = 30) did not express Reg IV. This is the first immunohistochemical analysis of the expression and distribution of Reg IV protein in human tumours. These data suggest that Reg IV is expressed by gastrointestinal and pancreatic tumours, including adenocarcinomas and carcinoid tumours, and that Reg IV is associated with intestinal and neuroendocrine differentiation of the stomach and gastric carcinoma.     

Reg IV expression is associated with cell growth and prognosis of adenoid cystic carcinoma in the salivary gland

Aims: Regenerating islet‐derived family, member 4 (Reg IV) is associated with the progression of various cancers. The aim was to examine Reg IV expression in adenoid cystic carcinomas (ACCs) in salivary glands. Methods and results: Reg IV expression was detected by immunohistochemistry and compared with clinicopathological parameters. Expression of phosphorylated epidermal growth factor receptor (pEGFR), phosphorylated AKT (pAKT) and MUC2 was examined by immunohistochemistry. Reg IV function was assessed with Reg IV antisense S‐oligodeoxynucleotides (AS) in ACC3 human ACC cells. Reg IV was expressed by salivary duct epithelia and acinus myoepithelia, but not in squamous epithelia. Reg IV expression was found in 41% (17/41) of ACCs, but in none of 40 oral squamous cell carcinomas (OSCCs) and was associated with nodal metastasis (P = 0.047) and poor prognosis (P = 0.012) in ACCs. Reg IV expression was associated with pEGFR (14/17, 82%) in Reg IV + ACCs, but had no relationship with pAKT or MUC2 expression in ACCs. Cell growth was inhibited by AS treatment in Reg IV + ACC3 cells, but not in HSC‐4 OSCC cell s , whereas in vitro invasion of neither cell types was affected by AS treatment. Conclusions: These results suggest that Reg IV might accelerate cell growth and disease progression of ACCs.     

Intestinally secreted C-type lectin Reg3b attenuates salmonellosis but not listeriosis in mice

The Reg3 protein family, including the human member designated pancreatitis-associated protein (PAP), consists of secreted proteins that contain a C-type lectin domain involved in carbohydrate binding. They are expressed by intestinal epithelial cells. Colonization of germ-free mice and intestinal infection with pathogens increase the expression of Reg3g and Reg3b in the murine ileum. Reg3g is directly bactericidal for gram-positive bacteria, but the exact role of Reg3b in bacterial infections is unknown. To investigate the possible protective role of Reg3b in intestinal infection, Reg3b knockout (Reg3b(-/-)) mice and wild-type (WT) mice were orally infected with gram-negative Salmonella enteritidis or gram-positive Listeria monocytogenes. At day 2 after oral Listeria infection and at day 4 after oral Salmonella infection, mice were sacrificed to collect intestinal and other tissues for pathogen quantification. Protein expression of Reg3b and Reg3g was determined in intestinal mucosal scrapings of infected and noninfected mice. In addition, ex vivo binding of ileal mucosal Reg3b to Listeria and Salmonella was investigated. Whereas recovery of Salmonella or Listeria from feces of Reg3b(-/-) mice did not differ from that from feces of WT mice, significantly higher numbers of viable Salmonella, but not Listeria, bacteria were recovered from the colon, mesenteric lymph nodes, spleen, and liver of the Reg3b(-/-) mice than from those of WT mice. Mucosal Reg3b binds to both bacterial pathogens and may interfere with their mode of action. Reg3b plays a protective role against intestinal translocation of the gram-negative bacterium S. enteritidis in mice but not against the gram-positive bacterium L. monocytogenes.     

Rapid determination of Epstein-Barr virus latent or lytic infection in single human cells using in situ hybridization.

Epstein-Barr (EBV) virus is associated with malignancies such as lymphoma and carcinoma. Infection of cells with EBV may result in either lytic infection with production of viral particles, characterized by the presence of linear DNA forms, or latent infection, characterized by either episomal or integrated DNA forms. To examine whether the different lytic and latent EBV DNA forms can reliably be distinguished in single human cells, in situ hybridization was performed in EBV-positive cell lines. Immunocytochemistry and Southern blot analysis were performed supplementary to in situ hybridization. In latent infection, three in situ hybridization patterns were observed: large-disperse (episomal), small-punctate (integrated) and combined (both), signal types 1, 2 and 3 respectively. These were associated with expression of latent membrane protein 1, but not with Z fragment of Epstein-Barr replication activator or viral capsid antigen. In lytic infection, three additional in situ hybridization patterns were observed: nuclear membrane associated, bubble (filling up the nucleus) and spillover (covering the lysed cells) signals types 4, 5 and 6 respectively. Signal types 4 and 5 were associated with expression of latent membrane protein 1 and Z fragment of Epstein-Barr replication activator but not viral capsid antigen, whereas type 6 was associated with expression of viral capsid antigen only. Southern blot analysis confirmed these results; however, low copy numbers of integrated virus were often missed by Southern blot, confirming that in situ hybridization is more sensitive in determining the presence of all types of EBV DNA. In situ hybridization may prove useful in rapidly screening large series of tissue microarrays and other clinical specimens for the presence of lytic or latent EBV.     

胰岛再生源蛋白与肝脏疾病和肝再生

胰岛再生源蛋白 (Reg) 是相对分子质量为16kD的分泌型蛋白,属于C类凝集素超家族成员。Reg蛋白是1个多功能分子,主要参与调控胰腺、胃、肠和肝脏中细胞的生长和增殖,在消化性疾病及癌症的发展和预后中发挥重要作用。我们主要综述了Reg对肝脏疾病和肝再生调控的研究进展,包含其在消化性疾病中介导的信号通路,为开展Reg在相关疾病诊疗及促进肝再生中的应用研究提供重要理论基础。     

RegⅠ在胃泌素促胃癌细胞体外增殖通路中的作用

目的 探讨再生蛋白I (RegⅠ)在胃泌素刺激胃癌细胞增殖通路中的作用.方法 根据RegⅠ cDNA已知序列,在线设计3个小干扰RNA,并经基因库同源性比较,构建其RegⅠ-shRNA表达载体(pSUPER-EGFP-REG/1、pSUPER-EGFP-REG/2和pSUPER-EGFP-REG/3),利用脂质体2000转染试剂分别转染胃癌细胞BGC823细胞株、SGC7901细胞株,同时设转染空质粒的实验对照组和无质粒转染的空白细胞组.后经G418筛选建立稳定转染细胞株.RT-PCR检测RegⅠ基因mRNA的转录水平,四甲基偶氮唑盐(MTT)法检测胃泌素孵育对胃癌细胞增殖的效应.结果 酶切及测序鉴定,插入片段正确,3个RegⅠ-shRNA表达载体构建成功;RT-PCR显示,pSUPER-EGFP-REG/1可有效抑制RegⅠ mRNA在胃癌细胞BGC823、SGC7901的转录.Western boltting结果显示,BGC823和SGC7901细胞的RegⅠ蛋白表达率分别下调到空载体对照组的(45±4)%和(53±4)%;MTT结果显示,RegⅠ基因表达抑制胃癌细胞系的增殖效率均降低(P＜0.05).胃泌素孵育后,胃癌细胞BGC823、SGC7901的增殖效率均增加(P＜0.05),而RegⅠ基因表达抑制的胃癌细胞系增殖效率则无明显改变(P＞0.05).结论 实验成功建立了RegⅠ基因表达抑制的胃癌细胞系; RegⅠ基因可能对胃泌素促胃癌细胞的增殖有促进的作用.