Peritoneal fibrosis (PF) represents a well-recognized complication associated with continuous ambulatory peritoneal dialysis therapy, characterized by a reversible epithelial-to-mesenchymal transition (EMT) at the early stage. The aim of the current study was to investigate the effects linked with the long noncoding RNA (lncRNA) AK089579 on the EMT of peritoneal mesothelial cells (PMCs) as well as the associated regulatory mechanisms of AK089579 downstream of tyrosine kinase 2 (DOK2) and microRNA-296-3p (miR-296-3p). Enrichment analysis, gene intersection association analysis, and a gene-gene intersection network were initially constructed to ascertain whether AK089579 regulated the expression of DOK2 through the mediation of miR-296-3p via the Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in PF. After the PF mouse model had been constructed, the expression of the proteins associated with the JAK2/STAT3 signaling pathway and EMT and PMC migration and invasion were all determined accordingly. Based on the obtained results, AK089579 was determined to function as a competing endogenous RNA for miR-296-3p while acting to up-regulate the expression of DOK2, which is a target gene of miR-296-3p. AK089579 was detected to confer an inhibitory effect on the activation of the JAK2/STAT3 signaling pathway, whereby the migration and invasion of PMCs among the mice models were suppressed. Meanwhile, up-regulated miR-296-3p and down-regulated DOK2 produced contrasting effects when compared with the aforementioned findings. Treatment with wp10066, a JAK2/STAS3 signaling pathway inhibitor, was shown to reverse the effects exerted by up-regulated miR-296-3p. Taken together, the central findings of the current study present evidence highlighting the capability of the lncRNA AK089579 to bind competitively to miR-296-3p and indirectly enhance the expression of DOK2, which in turn suppresses the activation of the JAK2/STAT3 signaling pathway, whereby the EMT, migration, and invasion of PMCs was inhibited in PF.

Effect of interference with chromatin remodeling and spacing factor-1 (RSF-1) on proliferation and apoptosis of breast cancer cells was investigated. MCF-7 and SKBR-3 cells were cultured in vitro and were divided into 3 groups: control group, negative siRNA control group (NC) and RSF-1 siRNA group. Western blot analysis was used to detect the expression of RSF protein after interference. Cell Counting Kit-8 (CCK-8) method was used to detect the effect of RSF-1 siRNA on cell proliferation. Plate cloning assay was used to detect the effect of RSF-1 siRNA on cell clone formation ability. Annexin V/PI double staining method was used to detect the effect of RSF-1 siRNA on cell apoptosis. Effect of RSF-1 siRNA on nuclear factor-κB (NF-κB) and its downstream signaling pathway were detected by western blot analysis. Western blot analysis showed that RSF-1 siRNA significantly downregulated the expression of RSF-1 protein in MCF-7 and SKBR-3 cells at 72 h after transfection (P<0.01). Cell proliferation assay showed that RSF-1 siRNA significantly reduced the proliferation ability and clone formation ability of MCF-7 and SKBR-3 cells compared with the control group (P<0.01). Annexin V/PI double staining assay results showed that compared with the control group, RSF-1 siRNA significantly increased the apoptosis rate of MCF-7 and SKBR-3 cells (P<0.01). Helenalin and Rsf-1 siRNA significantly reduced the expression levels of p-p65, Bcl-2, and XIAP proteins (P<0.01). Interfering with the expression of RSF-1, gene can effectively inhibit the proliferation of MCF-7 and SKBR-3 cells and promote their apoptosis. RSF-1 can be used as a potential new therapeutic target for the treatment of breast cancer.

The present study aimed to assess the expression and prognostic significance of remodeling and spacing factor 1 (RSF1; HBXAP) in renal cell carcinoma (RCC). RSF1 expression was analyzed using immunohistochemistry on tissue samples from a consecutive series of 137 patients with RCC who underwent tumor resection between November 2000 and March 2004. The associations between RSF1 expression, clinicopathological factors and patient survival were investigated. Immunohistochemistry revealed that RSF1 was highly expressed in 43.1% (59/137) of the RCC samples. RSF1 expression levels were associated with the T stage of the Tumor-Node-Metastasis grading system. Kaplan-Meier survival analysis indicated that high RSF1 expression in RCC was significantly associated with a poor prognosis. Multivariate analysis revealed that RSF1 expression is an independent prognostic parameter for the duration of overall survival of patients with RCC. The results demonstrated that a high expression level of RSF1 in RCC is associated with advanced tumor stages and a poor prognosis. To the best of our knowledge, the present study provides novel evidence of the biological significance of RSF1 expression in RCC.

Rsf-1 (HBXAP) was recently reported to play roles in tumorigenesis and tumor progression. There have been many reports referred to Rsf-1 overexpression in various cancers and associated with the malignant behavior of cancer cells. However, the molecular mechanism of Rsf-1 in non-small cell lung cancer aggressiveness remains ambiguous. In the present study, we found that there was a significant association between Rsf-1 overexpression and poor overall survival (p = 0.028) in lung cancer. Furthermore, knockdown of Rsf-1 expression in H1299 and H460 cells with high endogenous Rsf-1 expression inhibited cell migration and invasion and downregulated MMP2 expression and nuclear levels of NF-κB. NF-κB inhibitor could also block the effect of Rsf-1 in regulation of MMP2 expression. Further experiments demonstrated that Rsf-1 depletion restrained NF-κB reporter luciferase activity and downregulated bcl-2 and p-IκB protein level. In conclusion, we demonstrated that Rsf-1 was overexpressed in lung cancer and associated with poor survival. Rsf-1 regulated cell invasion through MMP2 and NF-κB pathway.

It remains controversial whether the addition of a second cytotoxic agent can further improve the therapeutic effect of gemcitabine monotherapy in advanced or metastatic pancreatic cancer (LA/MPC).

Objective:To evaluate the clinical correlation between PARP1 expression and triple-negative breast cancer.Methods:We searched PubMed,CNKI,Wanfang Database and other searches to collect literatures about the PARP1 expression in triple-negative breast cancer.Statistical analyses were carried out by using RevMan 5 .2 soft-ware.Results:Five literatures of eligible studies involving 700 cases,which including 331 triple -negative breast cancer patients and 369 non triple-negative breast cancer patients,met the inclusion criteria.Results of the Meta-a-nalysis showed that the expression of PARP1 in triple-negative breast cancer was significantly different from that of non triple-negative breast cancer (OR=3.24,95%CI:1.45 ~7.23,P=0.004).Conclusion:The expression of PARP1 was positively correlated with triple -negative breast cancer.It may be used as an indication to judge the prognosis of triple-negative breast cancer patients,but it still needs a large sample of randomized controlled trials for further study.Triple-negative breast cancer patients with conventional detection of PARP1 expression has some clini-cal significance.

目的:探讨6例肺滑膜肉瘤(primary pulmonary synovial sarcoma,PPSS)的病理特征及临床治疗情况.方法:HE染色后观察癌组织病理形态特征,SP免疫组化法检测其免疫表型.总结本病的临床,病理资料及诊治.结果:肺滑膜肉瘤的病理组织学特点具有向间叶、上皮组织双向分化的特征,6例中5例为梭形细胞型单相型滑膜肉瘤,1例为双相型滑膜肉瘤.肿瘤细胞形态差异小,紧密弥漫排列或交织束状排列,瘤细胞间可有少量胶原纤维,肿瘤细胞周围可见大小不一、扩张的裂隙,核分裂像多见.免疫组化结果显示,肿瘤细胞CK、CD99、Bcl-2、Vimentin及Ki-67阳性表达,TTF-1、CD34等阴性表达.肺滑膜肉瘤临床症状不特异,6例中4例均行一侧肺叶切除术,另外2例为会诊病例.结论:肺滑膜肉瘤是一种少见的肺部恶性肿瘤,恶性程度高.因其临床症状不特异,诊断主要依靠病理及免疫组化检查,分子遗传学检测对其确诊有意义.早期诊断及积极的手术治疗可能有效的改善其预后,提高患者生存率.

Objective To detect the expression of TRIM24 in uroepithelium cell carcinoma of bladder(BUCC)and analyze the relationship be?tween its expression and clinical pathological factors. Methods Immunohistochemical methods were employed to evaluate protein level of TRIM24 in BUCC. Results TRIM24 expression was observed in the nuclear compartments of tumor cells,while normal epithelia of bladder tissues exhibited negative results. For 72 BUCC tissues,TRIM24 overexpression(score≥4)was observed in 41.6%of cases(30/72). TRIM24 expression correlat?ed with invasive depth(P=0.029)and patients with high TRIM24 expression had high grade of BUCC(P=0.043). Conclusion There was sig?nificant over expression of TRIM24 in BUCC,which was correlated with poor differentiation and high grade of BUCC. It may become a candidate tar?get gene for the treatment of human uroepithelium cell carcinoma of bladder.

Objective:To study the expression of Rsf -1 in hepatocellular carcinoma and the relationship between its expression and clinical pathological factors.Methods:Immunohistochemical method was employed to evaluate pro-tein levels of Rsf -1 in hepatocellular carcinoma.Results:Rsf -1 expression was observed in the nuclear compart-ments of tumor cells,while normal liver tissues exhibited negative or weak expression.For 65 hepatocellular carcinoma tissues,negative and weak Rsf -1 expression (score <3)was observed in 56.9% (37 /65)of cases,while Rsf -1 overexpression (score ≥3)was observed in 43.1% cases (28 /65).There was no statistical difference between Rsf-1 overexpression and age (P =0.526),gender (P =0.230),hepatitis (P =0.835),cirrhosis (P =0.447),and nodal status (P =0.942).Strong Rsf -1 expression correlated with high serum AFP levels(P =0.026)and patients with high Rsf -1 expression had advanced stage of hepatocellular carcinoma (P =0.041).Conclusion:There was ex-pression of Rsf -1 in hepatocellular carcinoma.Strong Rsf -1 expression correlated with serum AFP levels and pa-tients with high Rsf -1 expression had advanced stage of hepatocellular carcinoma.

Nemo-like kinase (NLK), as a mitogen activated protein kinase (MAPK)-like kinase, is involved in the development of several human cancers. In this study, we explored the expression of NLK in lung squamous cell carcinoma (SCC) and adenocarcinoma tissues, and investigated the associations among NLK, β-catenin, T-cell factor 4 (TCF4), and the clinicopathological factors of lung cancers. The expressions of NLK, β-catenin, TCF4 were examined in 109 cases of lung cancers using immunohistochemistry method. The expression of NLK was observed in the nuclei of lung cancer tissues, and was significantly higher in lung cancer tissues than that in corresponding normal lung tissues (t = 21.636, n = 109, P < 0.001). The high expression of NLK was found in 45 cases of lung SCCs (45/49, 91.84%), which was much more than that in adenocarcinomas (38/60, 63.33%) (P = 0.001). Furthermore, the high expression of NLK was negatively correlated with TCF4 expression and positively correlated with the membranous expression of β-catenin. In conclusion, the present study demonstrated that the expression of NLK was localized in nucleus and significantly increased in lung cancers. The expression of NLK was negatively correlated with TCF4 expression and positively correlated with β-catenin membranous expression in lung cancers.

Tripartite motif-containing 24 (TRIM24), also known as transcription intermediary factor 1-alpha (TIF1α), is a chromatin-associated protein which as been has been implicated in carcinogenesis. However, its expression profile and biological roles in human bladder carcinoma has not been investigated. In this study, we examined its expression in 95 bladder cancer specimens. We found that TRIM24 expression was upregulated in 39 of 95 (41.1 %) specimens compared with normal control. TRIM24 overexpression was associated with local invasion and advanced grade of bladder cancer. In addition, we transfected TRIM24 plasmid into BIU-87 cell line and TRIM24 siRNA into 5637 cell line. Colony formation, CCK-8, and transwell assay were used to assess its biological roles in bladder cancer cells. The result showed that TRIM24 could facilitate cancer cell growth and invading ability. Western blot analysis demonstrated that TRIM24 upregulated cyclin D1, cyclin E, p-IκBα, and p-AKT expression, suggesting TRIM24 activates NF-κB and AKT pathways. In addition, NF-κB inhibitor reversed the effect of TRIM24 on cyclin D1. In conclusion, TRIM24 is overexpressed in human bladder cancer and facilitates bladder cancer growth and invasion, possibly through NF-κB and AKT signaling pathways.

目的:分析Tenascin-C(TN-C)在肾透明细胞癌中的表达及其与临床病理因素间的关系.方法:应用免疫组化方法检测肾透明细胞癌中TN-C蛋白表达情况.结果:免疫组化结果显示40.63％ (39/96)的病例存在肿瘤间质TN-C蛋白的过表达,且过表达与肿瘤组织核分级负相关(P=0.022);18.75％(18/96)的病例存在肿瘤细胞胞质TN-C蛋白的过表达,且过表达与肿瘤组织核分级、p-TNM分期正相关,分别为(P ＜0.001;P =0.038).肿瘤间质及肿瘤细胞胞质的过表达与RCCC患者的年龄、性别、肿瘤大小均无明显相关性.结论:肾透明细胞癌中存在TN-C蛋白肿瘤间质及肿瘤细胞的过表达,并与肿瘤分化和p-TNM分期相关.

Identifying patients at high risk of metastasis is a major challenge in lung adenocarcinoma (ADC) therapy, therefore discovery of noninvasive biomarkers and therapeutic targets is urgent. We found significant differences between the secretomes of differentially expressed proteins in lung ADC cell lines, clinical tissue samples and serum plasma samples with high and low metastatic potential. In particular, Apolipoprotein E (APOE) levels were three-times greater in cells with lymph node metastases (LNM) than those without. Our study indicates that APOE is a potential indicator of metastatic lung ADC and that secretomes may offer a valuable resource for biomarkers of lung ADC with LNM.

The aim of this study was to investigate c-Myc and -catenin-mediated drug resistance in A549/DDP lung adenocarcinoma cells. Cisplatin sensitivity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) toxicity assay. -Catenin and c-Myc protein expression following cisplatin treatment were determined using western blotting and immunofluorescence. Flow cytometry was performed to detect cell cycle and apoptosis in A549, A549/DDP, and c-Myc small interfering RNA (siRNA)-transfected A549/DDP cells before and after treatment with different doses of cisplatin. The median inhibitory concentration (IC50) in cisplatin-treated A549 and A549/DDP cells was 5.769 +/- 0.24mol/L and 28.373 +/- 0.96mol/L, respectively; the cisplatin resistance of A549 cells was about five times that of A549/DDP cells. Endogenous -catenin and c-Myc expression in A549/DDP cells were higher than that in A549 cells, and were upregulated in A549/DDP cells (p<0.05) and downregulated in A549 cells after 48h cisplatin treatment (p<0.05). -catenin localization transferred from membrane/cytoplasmic/nuclear to cytoplasmic/nuclear, and c-Myc localization transferred from cytoplasmic/nuclear to nuclear in both cell lines following cisplatin treatment. The rate of apoptosis increased in a dose-dependent manner with cisplatin. After 48-h transfection with c-myc siRNA, A549/DDP cells were blocked in the S phase, and G0/G1-phase cells increased. Simultaneously, the apoptotic rate was increased (p<0.05) and the IC50 decreased significantly (p<0.05). C-myc, the downstream target gene of -catenin, plays an important role in regulating cisplatin resistance in A549/DDP cells. C-Myc siRNA improved the sensitivity of A549/DDP cells to cisplatin.

目的:分析Rsf-1在非小细胞肺癌中的表达及与临床病理因素的关系.方法:应用Real-time PCR和免疫组化法检测非小细胞肺癌中Rsf-1的mRNA及蛋白表达情况.结果:有84％(21/25)肺癌组织病例中Rsf-1的mRNA含量明显高于对应的正常肺组织(P＜0.001),肿瘤组织中Rsf-1的mRNA平均值是其癌旁正常肺组织均值的2.37倍.免疫组化结果显示68.4％ (67/98)病例存在Rsf-1蛋白过表达,其过表达与肺癌患者的年龄、性别、肿瘤类型、原发肿瘤分级和淋巴结转移等未发现明显的相关性,而与肺癌的低分化和高p-TNM分期呈正相关(P=0.001;P=0.020).结论:肺癌中存在Rsf-1基因的扩增和蛋白的高表达,并与低分化和高p-TNM分期相关.

目的 检测染色质重塑因子1(Rsf-1)对肺癌增殖能力的影响,初步探讨Rsf-1促进肺癌增殖的分子机制.方法 应用Western blot方法筛选Rsf-1高表达人肺癌H1299、H460细胞系,通过小干扰RNA (siRNA)方法干扰内源性Rsf-1表达,并检测干扰效率.应用集落形成、流式细胞术检测Rsf-1干扰前后肺癌细胞增殖能力的变化.应用Western blot检测干扰内源性Rsf-1表达前后肺癌细胞中增殖相关因子的变化.结果 Western blot结果显示:肺癌细胞中Rsf-1蛋白表达水平明显高于正常支气管上皮HBE细胞系,其中H1299、H460细胞中存在非常明显的Rsf-1高表达.转染Rsf-1特异的siRNA于高表达Rsf-1的H1299和H460细胞后,其克隆形成明显少于对照组(H1299细胞中对照组和干扰组分别为123±15和83±9; H460细胞中对照组和干扰组分别为218±18和112±17)(P＜0.05).流式细胞术结果显示:干扰内源性Rsf-1表达后,H1299和H460细胞中Gt期细胞比例增多(H1299细胞中对照组和干扰组分别为56％±5％和71％±7％;H460细胞中对照组和干扰组分别为53％±4％和70％±6％),S期细胞比例减少(H1299细胞中对照组和干扰组分别为17％±2％和11％±5％;H460细胞中对照组和干扰组分别为19％±2％和10％±5％).细胞周期相关蛋白检测结果:干扰内源性Rsf-1表达后,H1299和H460细胞中cyclinD1和磷酸化细胞外调节蛋白激酶(pERK)水平明显下调.干扰Rsf-1在下调H1299和H460细胞pERK蛋白表达的作用结果与应用细胞外调节蛋白激酶(ERK)抑制剂U0126相似.结论 Rsf-1可通过cyclinD1/ERK相关通路促进肺癌细胞的增殖.

目的:探讨Wnt/β-catenin信号转导通路在肺腺癌顺铂耐药中的分子作用机制。方法:体外常规培养人肺腺癌A549细胞和其顺铂耐药A549/DDP细胞,取对数生长期的细胞用于实验。采用MTT法检测A549和A549/DDP细胞对顺铂的IC50;采用AnnexinⅤ/PI法和Hoechst 33342染色法检测顺铂处理后2种细胞的凋亡率;采用蛋白质印迹法检测2种细胞中β-catenin和Survivin的表达;采用siRNA干扰沉默β-catenin的表达,检测A549/DDP细胞的IC50值、细胞凋亡率及β-catenin和Survivin的表达。结果:顺铂对A549/DDP细胞的IC50值为（28.984±1.404）μmol/L高于A549细胞的（5.888±0.338）μmol/L,t=27.696,P＜0.001;20μmol/L顺铂诱导A549/DDP细胞凋亡率为（21.75±0.96）%,显著低于A549细胞的（39.38±0.88）%,t=23.474,P＜0.001。A549/DDP细胞中β-catenin蛋白的表达为1.890±0.060,显著高于A549细胞的1.063±0.035,t=20.595,P＜0.001;在A549/DDP细胞中Survivin蛋白表达量为1.107±0.061,明显高于A549细胞的0.503±0.025,t=15.814,P＜0.001。RNAi技术沉默β-catenin蛋白耐药性（14.615±0.939）μmol/L,显著低于未沉默β-catenin A549/DDP细胞的（28.984±1.404）μmol/L,t=14.732,P＜0.001;RNAi技术沉默β-catenin蛋白细胞凋亡率为（37.57±0.64）%,显著高于未沉默β-catenin A549/DDP细胞的（21.75±0.96）%,t=23.699,P＜0.001;RNAi技术沉默β-catenin蛋白下游靶基因Survivin蛋白的表达为0.527±0.065,显著低于A549/DDP组的1.027±0.025和瞬时转染阴性对照siRNA组的1.033±0.040,F=116.944,P＜0.001。结论:抑制Wnt/β-catenin信号转导通路的活性可以降低A549/DDP细胞对顺铂的耐药性。

The aim of this study was to investigate c-Myc and β-catenin–mediated drug resistance in A549/DDP lung adenocarcinoma cells. Cisplatin sensitivity was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) toxicity assay. β-Catenin and c-Myc protein expression following cisplatin treatment were determined using western blotting and immunofluorescence. Flow cytometry was performed to detect cell cycle and apoptosis in A549, A549/DDP, and c-Myc small interfering RNA (siRNA)–transfected A549/DDP cells before and after treatment with different doses of cisplatin. The median inhibitory concentration (IC 50 ) in cisplatin-treated A549 and A549/DDP cells was 5.769 ± 0.24 μmol/L and 28.373 ± 0.96 μmol/L, respectively; the cisplatin resistance of A549 cells was about five times that of A549/DDP cells. Endogenous β-catenin and c-Myc expression in A549/DDP cells were higher than that in A549 cells, and were upregulated in A549/DDP cells ( p  <   0.05) and downregulated in A549 cells after 48 h cisplatin treatment ( p  <   0.05). β-catenin localization transferred from membrane/cytoplasmic/nuclear to cytoplasmic/nuclear, and c-Myc localization transferred from cytoplasmic/nuclear to nuclear in both cell lines following cisplatin treatment. The rate of apoptosis increased in a dose-dependent manner with cisplatin. After 48-h transfection with c-myc siRNA, A549/DDP cells were blocked in the S phase, and G0/G1-phase cells increased. Simultaneously, the apoptotic rate was increased ( p  <   0.05) and the IC 50 decreased significantly ( p  <   0.05). C-myc, the downstream target gene of β-catenin, plays an important role in regulating cisplatin resistance in A549/DDP cells. C-Myc siRNA improved the sensitivity of A549/DDP cells to cisplatin.

目的:分析6例膀胱肉瘤样癌病理诊断、临床治疗情况及其随访结果.方法:HE染色后观察癌组织病理形态特征,SP免疫组化法检测其免疫表型.总结本病的临床及病理资料、诊治及预后.结果:病理结果显示6例膀胱肉瘤样癌均有不同级别的尿路上皮癌成分,同时伴有50％以上的肉瘤样成分.免疫组化结果显示,肉瘤样癌区域vimentin(VIM)及CK均阳性.治疗及预后情况显示,6例中3例行经尿道膀胱肿瘤切除术(TUR-BT),其中1例死亡,另外2例随访中;1例行膀胱部分切除术,术后间断行膀胱灌注化疗,之后失去随访;2例会诊病例资料不详.结论:膀胱肉瘤样癌是一种罕见的具有上皮性和间叶性双重分化特征的膀胱恶性肿瘤,其恶性程度高,预后不良.因其临床症状不特异,诊断主要依靠病理及免疫组化检查.早期诊断及积极的手术治疗能够有效的改善预后,提高患者生存率.

Objective To explore the role of GSK-3β in cisplatin resistance by evaluating the phosphorylation status and intracellular localization of GSK-3β in lung adenocarcinoma cell A549 and its cisplatin resistant cell clone A549/DDP. Methods Both total and nuclear/cytoplasmic GSK-3β, p-GSK-3βser9and p-GSK-3βtyr6expression were detected by Western blot. Cispatin resistance and cell apoptotic rate were measured by MTT and flow cytometry in both A549 and A549/DDP cells. Results Increased cytoplasmic p-GSK-3βser9 was observed in A549/DDP cells as compared to A549 cells(P < 0.01). Cisplatin increased cytoplasmic p-GSK-3βser9in A549/DDP cells but decreased it in A549 cells(P<0.01). Conversely, cytoplasmic levels of p-GSK-3βtyr216were significantly decreased in A549/DDP cells as compared to A549 cells. The levels of p-GSK-3βtyr216in the cytoplasm of A549/DDP cells decreased with cisplatin treatment while they increased in A549 cells(P<0.01). Conclusion Inhibition of cytoplasmic activity of GSK-3β was responsible for cisplatin resistance in lung adenocarcinoma.