目的 观察小干扰RNA (siRNA)沉默Kruppel样转录因子8(KLF8)基因对胃癌细胞MKN-28增殖及侵袭能力的影响.方法 构建针对KLF8的小干扰RNA(si-KLF8),应用实时定量聚合酶链反应(Real-time PCR)和Westem blot技术检测转染后胃癌细胞中KLF8 mRNA和蛋白的表达,应用噻唑蓝(MTT)比色法检测转染24、48、72 h的细胞增殖能力的变化,应用Transwell小室实验检测转染48 h后胃癌细胞侵袭能力的变化.结果 si-KLF8和阴性对照siRNA (si-CTRL)成功转染至胃癌细胞;si-KLF8组KLF8 mRNA和蛋白的表达量明显低于阴性对照组和正常对照组(P＜0.05);si-KLF8组增殖能力下降,24、48、72 h的吸光度值分别为0.125±0.003、0.193±0.005、0.223±0.005、0.267 ±0.003、0.315±0.006,穿膜细胞数为(42.0±3.6)个,均明显低于阴性对照组和正常对照组(P＜0.05);阴性对照组和正常对照组比较,上述指标差异无统计学意义(P＞0.05).结论 si-KLF8可抑制人胃癌细胞的增殖和侵袭能力,提示KLF8在胃癌的发生发展过程中发挥着重要的作用.

Objective To investigate the effects of small interference RNA (siRNA) targeting Kruppel like factor 8 (KLF8) gene (si-KLF8) on proliferation and invasion of gastric cancer cell MKN-28.Methods Synthesized siRNA targeting KLF8 was transfected into MKN-28,at the same time,gastric cancer cells transfected with negative siRNA were used as control.KLF8 mRNA and protein were detected by using real-time quantitative polymerase chain reaction (Real-time PCR) and Western blotting respectively at 24 h.The viability of cells was examined by using methyl thiazol tetrazolium (MTT) assay at 24,48 and 72 h.The cell invasion were assessed by Transwell assay.Results Si-KLF8 and negative control siRNA (si-CTRL) were transfected into gastric cancer cells successfully.KLF8 mRNA and protein in si-KLF8 group were lower than in negative control group and normal control group.The proliferations of cells were 0.125 ±0.003,0.193 ± 0.005,0.223 ±0.005,0.267 ± 0.003 and 0.315 ± 0.006 at 24,48 and 72

目的：探讨KLF8蛋白在膀胱癌中的表达，并分析其与临床病理特征及预后的关系。方法选择膀胱癌标本137例，正常膀胱组织标本19例，采用免疫组织化学方法，检测膀胱癌和正常组织中KLF8蛋白的表达，并结合临床资料进行分析。结果正常组织中KLF8蛋白的表达水平明显低于肿瘤组织（P＜0.001）。低级别和高级别膀胱癌中KLF8蛋白表达的阳性率分别为37.2%、58.5%，差异有统计学意义（P＜0.05）。非肌层浸润性膀胱癌和肌层浸润性膀胱癌中KLF8蛋白表达的阳性率44%和67.4%，差异有统计学意义（P＜0.05）。本组随访7~112个月，在肌层浸润性膀胱癌中，Kaplan- Meier生存显示：KLF8蛋白的阳性表达与膀胱癌患者的总生存率有关（P＜0.05）。结论 KLF8蛋白的表达在膀胱癌组织中明显高于正常组织，KLF8蛋白的表达与膀胱癌的分级、分期、预后有关；与患者的年龄、性别、肿瘤的数目无相关性（P＞0.05）。KLF8蛋白的检测有助于膀胱癌的诊断及预后的评估。

Objective To investigate the expression of KLF8 in bladder cancer and its relation to the prognosis of patients. Methods The expression of KLF8 in 137 urothelial bladder cancer and 19 normal bladder tissue samples was detected by im-munohistochemistry, and its relation to clinicopathological features was analyzed. Results The positive rate of KLF8 protein was significantly higher in bladder cancer than that in normal bladder tissue (P<0.001). The positive rate of KLF8 protein was 37.2%in low grade bladder cancer and 58.5%in high grade bladder cancer (P<0.05). It was significantly lower in non- muscle- invasive bladder cancer than that in muscle invasive bladder cancer (44%, 67.4%, P<0.05 ). The patient was followed up for 7~112 months, Kaplan- Meier analysis showed that positive expression of KLF8 was significantly associated with overal survival of pa-tients with muscle- invasive- bladder cancer (P<0.05). Conclusion The expression of KLF8 protein is high in bladder cancer, which is co

目的 探讨胆囊癌组织中Krüppel样因子8(KLF8)蛋白的表达及其与胆囊癌患者临床病理特征和预后的关系.方法 回顾性分析2008年1月至2012年12月上海交通大学医学院附属新华医院收治的40例胆囊癌患者的临床资料,采用免疫组织化学染色和Western blot法检测40例胆囊癌患者癌组织及相应癌旁组织中KLF8蛋白的表达情况,分析其与胆囊癌患者临床病理因素及预后之间的关系.采用门诊及电话方式进行随访,随访时间截至2013年11月.计数资料比较采用Pearson x2检验、矫正x2检验和Fisher确切概率法,计量资料比较采用t检验.采用Kaplan-Meier法绘制生存曲线,Log-rank法检验总体生存率的差异.结果 KLF8蛋白的表达主要定位于细胞核.KLF8蛋白在胆囊癌患者癌组织及癌旁组织中的阳性表达率分别为77.5％ (31/40)和27.5％ (11/40),两者比较,差异有统计学意义(x2=6.580,P＜0.05).Western blot分析结果显示:KLF8蛋白在胆囊癌患者癌组织及癌旁组织中的相对表达量分别为0.67 ±0.03和0.22 ±0.12,两者比较,差异有统计学意义(t=8.660,p＜0.05).胆囊癌组织中KLF8蛋白的阳性表达率在胆囊癌患者的性别、年龄、淋巴结转移和胆囊结石方面比较,差异无统计学意义(x2=0.755,0.227,0.029,0.062,P＞0.05);而在肿

Objective To investigate the expression of Krüppel-like factor 8 (KLF8) protein in the gallbladder cancer tissues and its relationship with the clinical pathological features and prognosis of patients.Methods The clinical data of 40 patients with gallbladder cancer were analyzed retrospectively.All the patients were admitted at the Xinhua Hospital of the Shanghai Jiaotong University from January 2008 to December 2012.The expressions of KLF8 protein in the gallbladder and paracarcinoma tissues were examined by the immunohistochemistry and the Western blot.Patients were followed-up through outpatient examination and telephone interview until November 2013.Count data were analyzed using the Pearson x2 test and Fisher''s exact probability.Quantitative data were analyzed using the t test.Survival curve was generated using the Kaplan-Meier method,and the survival analysis was conducted using the Log-rank test.Results The expression of the KLF8 protein was predominantly detected in

目的微小RNA(microRNA,miRNA)-10b被证实可促进多种肿瘤细胞的生长及增强其侵袭能力。文中旨在研究miRNA-10b对人低转移肺癌细胞95-C的增殖及侵袭力的影响及其作用机制。方法用脂质体法将miRNA-10b真核表达质粒瞬时转染入95-C,实验设置空白对照组、阴性对照组和miRNA-10b质粒转染组,实时定量荧光PCR(Real-Time PCR,RT-PCR,)检测各组细胞miRNA-10b的表达及KLF4mRNA的表达,细胞增殖实验检测各组细胞的增殖情况,Transwell实验检测各组细胞侵袭能力,Western blot检测各组细胞KLF4蛋白的表达。结果转染后48h,RT-PCR检测,miRNA-10b质粒转染组miRNA-10b表达(1.61±0.12)较空白对照组(1.01±0.08)、阴性对照组(0.86±0.07)明显上调(P0.05),阴性对照组与空白对照组差异无统计学意义(P0.05)。生长曲线反映miRNA-10b质粒转染组细胞生长速度明显高于空白对照组和阴性对照组,差异有统计学意义(P0.05),空白对照组和阴性对照组比较,差异无统计学意义(P0.05);Transwell结果显示,miRNA-10b质粒转染组[(188.0±15.1)/高倍显微镜]较空白对照组[(151.0±11.3)/高倍显微镜]、阴性对照组[(136.0±10.8)/高倍显微镜]有更多的细胞迁移到Transwell膜的另一侧(P0.05),空白对照组与阴性对照组差异

Objective MiRNA-10b is an important member of the MiRNA family , which has been proven that miRNA-10b can promote the growth and invasion of a variety of tumor cells .The aim of this study was to to investigate the effect and mechanism of miR-NA-10b on proliferation and invasion of low metastasis of lung cancer cell line (95-C). Methods The recombinant of miRNA-10b was transfected into 95-C by lipofectin method .The experiment set up 3 groups:blank control group , negative control group and miRNA-10b expression plasmid transfected group .MiRNA-10b expression level and KLF4mRNA expression level were detected by real-time fluores-cence quantitative PCR ( RTFQ-PCR) .The cell proliferation was detected by cell proliferation assay .The invasive ability of cell was de-tected by Transwell experiment .The expression of KLF4 protein was assessed by Western blot . Results At 48 hours after transfection, compared with blank control group (1.01 ±0.08) and negative control group (0.86 ±