miR-124抑制食管鳞癌细胞TE-1增殖及侵袭的研究

目的 分析微RNA-124(microRNA-124,miR-124)在食管癌组织中的表达情况,并探讨miR-124在食管鳞癌增殖及侵袭中的作用及机制.方法 采用实时荧光定量RT-PCR检测miR-124在18例食管癌组织(Ⅰ期6例,Ⅱ期8例,Ⅲ期4例,Ⅳ期0例)及相应的癌旁组织中的表达情况.在食管癌细胞TE-1中转染miR-124 mimic,采用细胞集落形成实验、细胞计数试剂盒-8(cell counting kit-8,CCK-8)、细胞划痕实验检测miR-124过表达对食管癌细胞增殖及侵袭的影响.通过Targetscan预测细胞周期蛋白依赖性激酶(cyclin-dependent kinases 4,CDK4)为miR-124的直接靶基因,并用荧光素酶报告基因实验及蛋白质印迹法实验进行验证.结果 食管癌组织miR-124的表达水平较癌旁正常组织明显下降(P＜0.01).通过转染miR-124 mimics使TE-1细胞过表达miR-124,过表达miR-124后TE-1细胞增殖及侵袭能力明显下降(P＜0.01).结论 miR-124在食管癌组织中低表达,miR-124可能通过调控CDK4表达来影响食管癌细胞TE-1增殖及侵袭能力.     

miR-124 通过靶向BECN1 基因调控细胞自噬抑制食管癌KYSE170 细胞的侵袭和迁移

目的：探讨miR-124 通过调控细胞自噬对食管癌KYSE170 细胞侵袭和迁移能力的影响。方法：食管癌KYSE170 细胞转染miR-124 mimic,Transwell 实验检测细胞侵袭和迁移能力的变化,双荧光素酶报告基因验证miR-124 对BECN1(Beclin1)基因的靶向调控作用,Western blotting 分析对BECN1、P62 及LC3 蛋白表达水平的影响。向KYSE170 细胞中转染BECN1 siRNA沉默BECN1 的表达,Transwell 法检测细胞侵袭及迁移能力的变化,Western blotting 检测BECN1、P62 及LC3 蛋白的表达。将miR-124 mimic 与BECN1 过表达质粒共转染至KYSE170 细胞,Transwell 实验检测细胞侵袭及迁移能力的变化,Western blotting 检测自噬相关基因表达的变化。结果：转染miR-124 mimic 后,KYSE170 细胞的侵袭和迁移能力下降(P<0.05),BECN1 蛋白及荧光素酶报告基因活性均明显下调(均P<0.01),自噬相关蛋白P62 表达增高,LC3 表达水平明显降低(均P<0.01)。沉默BECN1 表达抑制食管癌细胞侵袭及迁移(P<0.01),而过表达BECN1 使miR-124 mimic 对KYSE170 细胞自噬、侵袭和迁移能力的抑制作用明显减弱(P<0.01),自噬相关蛋白P62 表达降低、LC3 蛋白表达水平明显升高(均P<0.01)。结论：miR-124 能够抑制食管癌细胞侵袭及迁移能力,其机制可能与靶向调控自噬相关基因BECN1 的表达影响细胞自噬有关。     

钠氢交换蛋白1基因在体外对食管癌KYSE-70细胞增殖和侵袭作用的影响

目的:观察钠氢交换蛋白1(Na+/H-exchanger 1,NHE-1)基因对食管癌KYSE-70细胞增殖和侵袭能力的影响.方法:采用实时荧光定量PCR和蛋白质印迹法检测NHE-1 mRNA及其蛋白在人食管癌EC-9706、KYSE-450、EC-1和KYSE-70细胞中的表达情况.将化学合成的靶向NHE-1的小干扰RNA (small interference RNA,si RNA)体外转染入KYSE-70细胞,实时荧光定量-PCR和蛋白质印迹法检测转染24、48和72 h后KYSE-70细胞中NHE-1 mRNA及其蛋白的表达,MTT和Boyden小室法检测KYSE-70细胞的增殖及侵袭能力.结果:NHE-1 mRNA及其蛋白在KYSE-70细胞中高表达.与阴性对照(转染阴性对照siRNA)及空白对照(未转染的KYSE-70细胞)组相比,靶向NHE-1的siRNA可抑制KYSE-70细胞中NHE-1 mRNA及其蛋白的表达,差异具有统计学意义(P＜0.05);与阴性对照组、空白对照组及NHE-1 siRNA转染24 h组比较,NHE-1 siRNA转染48和72 h后的KYSE-70细胞体外增殖能力和侵袭能力明显受到抑制,差异有统计学意义(P＜0.05).结论:干扰NHE-1基因的表达可以抑制食管癌KYSE-70细胞的体外增殖和侵袭能力,NHE-1基因有望成为食管癌基因治疗的重要靶点.     

miR-302c在胃癌中表达及对胃癌细胞侵袭和迁移影响的机制研究

摘 要：［目的］ 探讨miR-302c对人胃癌细胞侵袭和迁移能力的影响及机制。［方法］ 实时荧光定量PCR法检测miR-302c在胃癌细胞系和正常胃黏膜上皮细胞中的表达情况。在SGC7901和AGS细胞中分别转染miR-302c mimic和inhibitor后,Tranwell实验检测细胞侵袭和迁移能力变化。实时荧光定量PCR和Western blot法检测转染后环磷酸腺苷反应元件结合蛋白1(CREB1)的变化情况。［结果］与永生化的正常胃黏膜上皮细胞GES-1相比,miR-302c在胃癌细胞系SGC7901、MKN28、N87、MKN45和AGS中表达降低。转染 mimic和inhibitor后,miR-302c在SGC7901和AGS中表达分别明显上调和下调(P＜0.001)。Transwell实验发现,上调miR-302c后,SGC7901细胞的侵袭和迁移能力明显降低,而下调miR-302c后,AGS细胞的侵袭和迁移能力明显增强。进一步研究发现,miR-302c可抑制CREB1的表达。功能挽救实验证明CREB1可部分恢复miR-302c上调对SGC7901细胞侵袭和迁移的抑制作用。［结论］ miR-302c抑制胃癌细胞的侵袭和迁移,其机制可能是通过直接靶向CREB1。     

miR-452-5p靶向SOX7促进食管癌KYSE-150细胞增殖、侵袭及EMT

目的：检测miR-452-5p在食管鳞状细胞癌（ESCC）中的表达,并探讨其异常表达对食管癌KYSE-150细胞增殖、侵袭能力和EMT进程的影响及其分子机制。方法：收集2012年3月至2015年12月在河北医科大学第四医院就诊的86名ESCC患者的癌组织样本和对应的癌旁组织,用qPCR法检测miR-452-5p及其他相关基因在ESCC组织和细胞中的表达;向KYSE-150细胞中分别转染miR-452-5p mimic或pcDNA3.1-SOX7构建过表达的细胞株。分析miR-452-5p表达与ESCC病理特征和患者5年OS的关系。用MTS、Tanswell法检测miR-452-5p过表达对食管癌KYSE-150细胞增殖、侵袭能力和EMT进程的影响;用双荧光素酶报告基因实验及TOP/FOP报告基因系统检测miR-452-5p与SRY盒转录因子（SOX7）3’UTR区的结合作用及对Wnt/β-catenin通路活化水平的影响。结果：miR-452-5p在ESCC组织中呈明显高表达（P<0.01）,并与ESCC患者的淋巴结转移、TNM分期及5年OS密切相关（均P<0.01）。mi...     

MicroRNA-506靶向Caveolin-1抑制绒毛膜癌细胞增殖和侵袭的研究

摘 要：［目的］ 探讨microRNA-506 (miR-506)在绒毛膜癌中对绒毛膜癌细胞的作用及其作用机制。［方法］ 通过qRT-PCR检测绒毛膜癌和正常早孕绒毛临床标本中miR-506及caveolin-1表达量及相关性;将绒毛膜癌细胞JEG3和JAR按照miR-506 mimic（mimic）、miR-506 inhibitor（Inhi）、miR-506 mimic negative control（NC）和空白对照组（Blank）分别进行转染,通过MTT实验、Transwell实验和流式细胞术检测miR-506对绒毛膜细胞作用;通过双荧光素酶报告实验、qRT-PCR和Western blot检测miR-506、caveolin-1及相关蛋白的表达变化。［结果］ 与正常早孕绒毛相比,miR-506在绒毛膜癌组中表达显著降低（P<0.05）,并与caveolin-1表达呈负相关。MiR-506 过表达明显抑制绒毛膜癌细胞的增殖、侵袭并促进凋亡。双荧光素酶报告实验表明miR-506过表达显著抑制野生型荧光素酶活性,qRT-PCR和Western blot结果显示miR-506过表达显著抑制caveolin-1表达,进而抑制相关蛋白表达。［结论］ MiR-506可能通过靶向caveolin-1抑制绒毛膜癌细胞的增殖和侵袭。     

miR-124-3p.1靶向PRDM2调节JAK/STAT通路对胰腺癌细胞增殖和凋亡的影响

摘 要：［目的］ 研究miR-124-3p.1对胰腺癌细胞增殖、凋亡的影响,并探讨其作用机制。［方法］ 运用qRT-PCR法检测组织和细胞中miR-124-3p.1、PR结构域蛋白2(PRDM2)的表达;将miR-124-3p.1 mimics、miR-NC、anti-miR-124-3p.1 mimics、anti-miR-NC用脂质体法转染PANC-1细胞;Western blot检测细胞中PRDM2、p-JAK2、p-STAT3蛋白表达;MTT法检测细胞增殖。［结果］ 与人正常胰腺组织细胞相比,胰腺癌组织细胞中PRDM2表达显著性升高,miR-124-3p.1表达显著性降低（P<0.05）;PRDM2为miR-124-3p.1的靶标。过表达miR-124-3p.1可抑制胰腺癌细胞增殖、促进凋亡、抑制JAK/STAT通路。［结论］ miR-124-3p.1可抑制胰腺癌细胞增殖和促进凋亡,可能与靶向PRDM2和抑制JAK/STAT通路有关。     

miRNA-26b-3p通过靶向STAT3调控食管鳞状细胞癌细胞的增殖和迁移

目的：观察miR-26b-3p在食管鳞状细胞癌（ESCC）组织中的表达水平及其对ESCC细胞增殖、侵袭和迁移能力的影响,并探讨其分子调控机制。方法: 选取河北医科大学第四医院2018年4月1日至2018年12月25日手术切除的ESCC组织及相应癌旁组织各60例,利用qPCR法检测ESCC组织、癌旁组织和ESCC细胞中miR-26b-3p的表达。选取miR-26b-3p表达水平较低的ESCC细胞TE1和KYSE150,转染miR-26b-3p mimic,并设立阴性对照。利用细胞增殖实验、划痕愈合实验和Transwell实验检测过表达miR-26b-3p对TE1和KYSE150细胞增殖、迁移和侵袭能力的影响。荧光素酶报告基因实验检测miR-26b-3p与STAT3的 3''UTR 靶点部位的结合情况。随后同时转染 miR-26b-3p mimic 和 pcDNA3.0-STAT3,利用细胞增殖实验、划痕愈合实验和Transwell实验检测STAT3是否可逆转过表达miR-26b-3p对细胞增殖、迁移和侵袭能力的影响。qPCR 和 WB法检测甲基化酶抑制剂5-Aza-DC对ESCC细胞甲基化的影响和miR-26b-3p与STAT3表达的影响。结果: miR-26b-3p在ESCC组织中的表达低于癌旁组织（P<0.01）,其在 ESCC 细胞TE1、KYSE150和LYSE170中表达水平显著低于人正常食管上皮细胞HEEC（均P<0.01）。与miR-26b-3p NC组相比,miR-26b-3p mimic转染可明显上调TE1和KYSE150细胞中miR-26b-3p的表达（均P<0.01）,但可明显抑制两种细胞的增殖、迁移和侵袭能力（P<0.05或P<0.01）。荧光素酶报告基因实验结果表明,在TE1和KYSE150细胞中,miR-26b-3p明显抑制了野生型STAT3载体的荧光素酶活性（P<0.05或P<0.01）,而突变型的荧光素酶活性不受影响。同时转染miR-26b-3pmimic和pcDNA3.0-STAT3可部分逆转miR-26b-3p mimic对TE1 和 KYSE150 细胞增殖、迁移和侵袭能力的抑制作用（P<0.05或P<0.01）。5-Aza-DC 处理后 ,TE1 和 KYSE150 细胞中 miR-26b-3p 表达上调（均 P<0.01）、STAT3 mRNA 和蛋白水平降低（P<0.05）,miR-26b-3p呈现去甲基化状态。结论: miR-26b-3p的启动子高甲基化导致其在ESCC组织和细胞中的表达下调,其作为抑癌因子可通过靶向STAT3而抑制ESCC细胞的增殖、侵袭和迁移能力。     

miR-198靶向MAPK1调控宫颈癌HeLa细胞增殖、凋亡和侵袭

目的 探究miR-198对宫颈癌细胞增殖、凋亡和侵袭的作用及其机制。方法 用miR-198 mimic转染HeLa细胞,qRT-PCR检测miR-198和丝裂原活化蛋白激酶1 （Mitogen-activated protein kinase1, MAPK1）的mRNA水平;荧光素酶实验检测miR-198和MAPK1的靶向关系;用pcDNA3.0-MAPK1（pc-MAPK1）和miR-198 mimic转染细胞,CCK-8检测细胞增殖活性,流式细胞术检测细胞凋亡,Transwell法检测细胞侵袭能力,Western blot检测蛋白的表达。结果 miR-198 mimic转染细胞后,miR-198表达水平明显升高,MAPK1 mRNA表达水平明显降低;荧光素酶报告实验表明miR-198序列上存在MAPK1结合位点;miR-198过表达能显著降低宫颈癌细胞增殖倍数、诱导细胞凋亡,同时还能明显降低HeLa细胞侵袭能力;此外,miR-198 mimic能显著抑制MAPK1下游基因核糖体S6激酶2（Ribosomal S6 kinase 2, RSK2）、c-Myc和c-fos的蛋白表达;pc-MAPK1能明显减弱miR-198 mimic对HeLa细胞增殖、凋亡、侵袭及对MAPK1下游蛋白表达的调控作用。结论 miR-198过表达能通过靶向MAPK1抑制宫颈癌细胞的增殖和侵袭能力并诱导细胞凋亡。     

Hsa-miR-132通过靶向FOXA1基因影响食管癌细胞的增殖和侵袭

目的:探讨hsa-miR-132(miR-132)对人食管癌细胞增殖和侵袭的影响及其可能的机制。方法:构建pCDNA3.1(+)-miR-132表达载体并转染高转移潜能的人食管癌细胞KYSE150,G418筛选稳定过表达miR-132的KYSE150细胞。软琼脂克隆形成实验和Transwell小室实验检测KYSE150细胞的增殖和侵袭,Real-time PCR和Western blotting检测miR-132和叉头框蛋白A1基因(forkhead box protens A1,FOXA1)在KYSE150细胞中的表达,双荧光素酶报告基因分析和Western blotting检测过表达miR-132对FOXA1的调控作用。结果:成功构建pCDNA3.1(+)-miR-132质粒,获得稳定过表达miR-132的KYSE150细胞。亲本KYSE150细胞中miR-132低表达,而FOXA1蛋白高表达。与转染空载体和未转染KYSE150细胞相比,转染pCDNA3.1(+)-miR-132不影响KYSE150细胞的增殖(13.9±0.33 vs 15.4±0.11、17.1±0.20,P>0.05),但细胞体积较小且表面比较光滑;其可显著降低KYSE150细胞的侵袭能力[(55±1.6)vs(129.0±3.1)、(124.0±2.8)个细胞,P<0.01]。miR-132能够作用于FOXA1 3’-UTR,从而抑制内源性FOXA1的表达。结论:食管癌KYSE150细胞低表达miR-132,外源性过表达miR-132可负向调控FOXA1的表达,从而抑制KYSE150细胞的侵袭能力。     

E-钙粘蛋白及PTEN基因编码蛋白与胃癌浸润转移

目的:观察抑癌基因PTEN蛋白和ECD在胃癌组织中的表达,探讨其与胃癌生物学行为及预后的关系。方法:以兔抗人PTEN多克隆抗体、鼠抗人ECD单克隆抗体,采用SABC免疫组化法,检测100例胃癌手术切除标本中拟测指标的表达。以χ2和Logrank检验对结果做统计学分析。结果:ECD、PTEN蛋白在非癌胃粘膜中均见表达;在胃癌组织中表达下调或缺失。ECD异常表达率为42.0%;弥漫型胃癌异常表达率(48.57%),明显高于肠型胃癌(26.67%),(P<0.05);ECD异常表达与浸润深度有关(P<0.05)。胃癌组织中PTEN蛋白缺失率为59%;弥漫型胃癌缺失率(65.71%)明显高于肠型胃癌(43.33%),(P<0.05);伴淋巴结转移的胃癌缺失率(64.47%)明显高于无淋巴结转移者(41.67%),(P<0.05);PTEN蛋白缺失的患者比阳性表达者预后差(P=0.0066)。65.85%PTEN阳性表达者同时伴ECD正常表达。结论:两种标志物与胃癌浸润转移有关,PTEN表达与胃癌患者预后密切相关。将两种指标联合检测,可作为正确判断胃癌患者预后,指导临床治疗的分子生物学指标。     

Cytologic classification of precancer and cancer of the endometrium and esophagus and of malignant lymphomas

The paper discusses cytological classifications of precancer and cancer of the endometrium, esophagus and malignant lymphomas presented by cytologists from five Soviet research institutes of oncology. The classifications were based on the data of 4400 cases in conformity with WHO histologic classifications.     

世界卫生组织骨质疏松症防治工作报告和防治建议

引 言 作为对第51号综合处理非传染性疾病预防与控制的世界卫生组织决议的反应,1998年7月WHO成立了致力于不断完善对骨质疏松预防和治疗策略的工作小组。小组成员来自世界各国致力于骨质疏松研究的知名专家。Harry K.Genant为本届主席。这一项世界范围内的骨质疏松教育计划旨在通过世界范围的研究,不断改善对骨质疏松的诊断水平和发展并完善对骨质疏松病人的合理治疗。其重点将以发展中国家为主。并为各国政府及其卫生部门和病人群体提供世界性有关骨质疏松症的总体的、完整的指导性资料。该项研究、教育计划的实施将由世界各国的骨质疏松症研究和治疗机构共同完成,并经权威学术机构、政府和非政府组织进行有针对性的回顾研究,最终由WHO审议通过。     

Schwannomas of Head and Neck and Review of Literature

Benign nerve cell tumours have been given various names like schwannoma, neurilemmoma, neurinoma, neurofibroma, spindle cell tumours etc. Extra cranial head and neck schwannomas usually present as solitary and well-demarcated lesions. The lesion can cause secondary symptoms, such as nasal obstruction, dysphasia, and hoarseness, depending upon the location of the lesion. Fine needle aspiration cytology, CT scans, and MRI may be of limited help in the diagnosis of schwannomas. The treatment is complete surgical excision of the benign tumour and postoperative histopathological examination establishes the final diagnosis.     

Quality of life and care in leukemia,myeloma and non-malignant disease. Opinions of patients and relatives,and effects of geography and time

In a questionnaire study 140 subjects answered 4200 questions in 1980 and 1986. They consisted of patients with myeloma, acute leukemia, lung carcinoma, and non-malignant disease and their relatives. In 22 additional cases the questionnaire was not answered. The results show that myeloma patients are less content with the general care than leukemia patients (P < 0.05). Similarly, relatives of deceased myeloma patients are less satisfied with the information given to them than relatives of deceased leukemia patients (P < 0.001). The information has improved with time, however, since the patients were more satisfied in 1986 than in 1980 (P < 0.001) and relatives of myeloma patients still alive were more satisfied than relatives of patients who had died earlier (P < 0.001).     

A comparative study of knowledge and awareness of colorectal and breast cancerA comparative study of knowledge and awareness of colorectal and breast cancer

Aims: To assess and compare knowledge and awareness of colorectal cancer and breast cancer in a sample of the general population. Methods: Eleven hundred visitors to six different outpatient clinics, in a University Hospital, were given a study-specific questionnaire, based on educational material from the British Association of Cancer United Patients (CancerBACUP). The questionnaire consisted of 12 statements on the incidence, presentation, detection, treatment and prognosis of colorectal and breast cancer. Results: One thousand and sixty-eight individuals returned the questionnaire. One thousand and four completed questionnaires were analysed. The mean age (SD) of respondents was 50.1 (17.2) years, and the male to female ratio was 2:3. Respondents had read more about breast than about colorectal cancer (60.3%vs 32.4%,P <0.0001, McNemar's test). The proportion of correct answers for each statement on breast cancer was higher than for answers to corresponding items on colorectal cancer. Mean overall scores (95% CI) for breast and colorectal cancer were 88.1 (86.9, 89.2) and 64.4 (62.5, 66.3) respectively, the mean difference (95% CI) being 23.7 (22.0, 25.5). Scores were higher for breast cancer irrespective of age or gender. Conclusion: There is a low level of understanding of colorectal cancer in the general population when compared to breast cancer. This highlights the importance of public education in this common cancer.     

Prospective study of fruit and vegetable consumption and incidence of colon and rectal cancers

BACKGROUND: Frequent consumption of fruit and vegetables has been associated with a reduced risk of colorectal cancer in many observational studies. METHODS: We prospectively investigated the association between fruit and vegetable consumption and the incidence of colon and rectal cancers in two large cohorts: the Nurses' Health Study (88 764 women) and the Health Professionals' Follow-up Study (47 325 men). Diet was assessed and cumulatively updated in 1980, 1984, 1986, and 1990 among women and in 1986 and 1990 among men. The incidence of cancer of the colon and rectum was ascertained up to June or January of 1996, respectively. Relative risk (RR) estimates were calculated with the use of pooled logistic regression models accounting for various potential confounders. All statistical tests were two-sided. RESULTS: With a follow-up including 1 743 645 person-years and 937 cases of colon cancer, we found little association of colon cancer incidence with fruit and vegetable consumption. For women and men combined, a difference in fruit and vegetable consumption of one additional serving per day was associated with a covariate-adjusted RR of 1.02 (95% confidence interval [CI] = 0.98-1.05). A difference in vegetable consumption of one additional serving per day was associated with an RR of 1.03 (95% CI = 0.97-1.09). Similar results were obtained for women and men considered separately. A difference in fruit consumption of one additional serving per day was associated with a covariate-adjusted RR for colon cancer of 0.96 (95% CI = 0.89-1.03) among women and 1. 08 (95% CI = 1.00-1.16) among men. For rectal cancer (total, 244 cases), a difference in fruit and vegetable consumption of one additional serving per day was associated with an RR of 1.02 (95% CI = 0.95-1.09) in men and women combined. None of these associations was modified by vitamin supplement use or smoking habits. CONCLUSIONS: Although fruits and vegetables may confer protection against some chronic diseases, their frequent consumption does not appear to confer protection from colon or rectal cancer.     

Quality of life and care in leukemia, myeloma and non-malignant disease. Opinions of patients and relatives, and effects of geography and time

In a questionnaire study 140 subjects answered 4200 questions in 1980 and 1986. They consisted of patients with myeloma, acute leukemia, lung carcinoma, and non-malignant disease and their relatives. In 22 additional cases the questionnaire was not answered. The results show that myeloma patients are less content with the general care than leukemia patients (P less than 0.05). Similarly, relatives of decreased myeloma patients are less satisfied with the information given to them than relatives of deceased leukemia patients (P less than 0.001). The information has improved with time, however, since the patients were more satisfied in 1986 than in 1980 (P less than 0.001) and relatives of myeloma patients still alive were more satisfied than relatives of patients who had died earlier (P less than 0.001). The opinions of patients were similar to those of their relatives. However, the relatives of leukemia patients were even more satisfied with the contact with the medical staff than the patients themselves (P less than 0.05). As many as 10-30% of the relatives never gave up hope for their relative's survival. Only two out of 27 deaths were considered not dignified. The lung carcinoma patients reported a less good quality of life (P less than 0.001), and less satisfaction with the information given (P less than 0.01), than the hematological patients from the same year. Similarly, their attitude to the medical care improved less (P less than 0.01), and they were less content with the general care than the leukemia group (P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)