肿瘤组织中具核梭杆菌丰度作为Ⅱ期结直肠癌预后标志物的临床研究

目的 评估肿瘤组织中具核梭杆菌(Fn)丰度对Ⅱ期结直肠癌(CRC)患者预后的影响。方法 收集2013年1月—2018年12月151例在扬州大学附属医院接受手术治疗的Ⅱ期CRC患者的肿瘤标本,通过定量聚合酶链反应检测肿瘤组织中Fn的基因相对表达水平,并通过中位数法将其分为Fn高丰度组和Fn低丰度组。采用卡方检验分析Fn丰度与临床特征的相关性。采用Kaplan-Meier生存曲线分析Fn丰度对患者肿瘤特异性生存期(CSS)和无病生存期(DFS)的影响。基于Cox风险比例回归模型的单因素和多因素明确Fn丰度是否为预后的独立影响因素。结果 Fn丰度与Ⅱ期CRC患者临床特征均无显著相关性(P>0.05)。Ⅱ期CRC患者中,Fn丰度与患者的CSS及DFS呈负相关(P=0.01)。单因素及多因素分析结果表明,Fn丰度为影响Ⅱ期CRC患者CSS及DFS的独立影响因素(P<0.05)。亚组分析结果表明,Fn丰度与Ⅱ期低危患者的CSS及DFS无显著相关性(P=0.22、0.47),但与高危患者的CSS及DFS呈负相关(P<0.01)。单因素及多因素分析结果表明,Fn丰度为Ⅱ期高危CRC患...     

Toll样受体4与结直肠癌的相关性

Toll样受体4(toll-like receptor 4,TLR4)在炎症诱发的结直肠癌(colorectal cancer,CRC)中的作用日益受到关注.活化的TLR4通过髓样分化因子88 (myeloid differentiation factor 88,MyD88)激活核转录因子-κB (nuclear factor-κB,NF-κB)产生大量炎症介质并募集炎症细胞形成肿瘤微环境,从而成为炎症与肿瘤的纽带贯穿于CRC发生、发展的整个过程中.本文着重对TLR4信号通路参与CRC的起始、进展、转移、遗传变异和表观遗传调控进行了讨论.尤其,文章中强调了TLR4在CRC发展和发病机制中的角色,并提出调节TLR4信号通路可能为CRC的治疗带来新的前景.     

肠道菌群及其代谢产物在结直肠癌中的研究进展

结直肠癌(colorectal cancer,CRC)约占全球所有新发癌症的10%。CRC的病因和发病机制尚不完全清楚。由大量肠道微生物组成的肠道菌群位于肠上皮附近,参与机体能量获取、新陈代谢和免疫反应等生理活动。越来越多的证据表明肠道菌群失调可通过直接或间接的方式改变宿主的生理过程来影响CRC的发生、发展和对治疗的反应。此外,肠道菌群影响CRC的机制可以产生新的诊断和治疗方法,但仍有许多未知之处。     

MiRNA在结直肠癌干细胞信号通路中的作用

MiRNA是一类内源性非编码的单链小RNA,可以通过调控结直肠癌干细胞(colorectal cancer stem cell,CCSC)等参与结直肠癌(colorectal cancer,CRC)发生发展的各种过程。CCSC被认为是导致CRC发生发展、转移复发以及耐药性的主要原因,mi RNA在CCSC中差异表达,并且可在CRC组织和不同体液中保持一定的特异性和稳定性,因而mi RNA可作为CRC临床早期检测诊断、疗效预测及个体化治疗的新型生物标志物。     

TWEAK/Fn14mRNA在大鼠骨性关节炎滑膜中表达的实验研究

目的:研究TWEAK/Fn14是否参与了骨性关节炎(osteoarthritis,OA)发生发展,进一步探讨骨性关节炎发生机制。方法:将24只成年SD大鼠随机分为3组,即正常组、模型组、TWEAK抗体组,每组8只。模型组、TWEAK抗体组均采用木瓜蛋白酶关节腔注射建立骨性关节炎模型。TWEAK抗体组在造模前1 h经鼠尾静脉注射TWEAK抗体进行干预。检测指标:(1)关节滑膜HE染色;(2)关节滑膜TWEAK和Fnl4 mRNA的表达;(3)IL-1β、TNF-α在滑膜中的表达。结果:(1)HE染色:TWEAK抗体组较模型组改变轻,存在部分增生;(2)与模型组比较,TWEAK抗体组TWEAK/Fn-14mRNA表达均降低,差异有统计学意义(P<0.05);(3)与模型组比较,TWEAK抗体组滑膜中IL-1β、TNF-α表达降低,差异有统计学意义(P<0.05)。结论:TWEAK/Fn14参与了OA的发生,可能是OA形成的机制之一。     

结直肠癌患者相关危险因素分析

目的探讨结直肠癌患者危险因素及其临床意义。方法回顾性分析重庆市第十三人民医院和重庆医科大学附属第二医院近10年400例结直肠癌(CRC)患者(病例组)临床资料,收集同期400例未合并肿瘤、消化系统疾病患者为对照组,分析两组性别、年龄、糖尿病(DM)发病率及病程、体质量指数、结直肠癌家族史的区别及相关性。结果 (1)病例组中男性多于女性,且随年龄增加,男性比例升高;(2)病例组中合并DM发病率为25.0%,明显高于对照组的12.0%,差异有统计学意义(P0.05);(3)CRC家族史、超重和肥胖、DM是CRC发生的独立危险因素;(4)CRC发生风险与DM病程呈正相关(P0.05),DM病程4年、4～8年及8年患者CRC发生风险分别为1.79(95%CI:1.05～3.06)、2.52(95%CI:1.52～4.20)、5.54(95%CI:3.52～8.73);(5)CRC合并DM患者为中晚期64.6%,明显高于CRC不合并DM患者的29.2%,差异有统计学意义(P0.05)。结论有CRC家族史、超重及肥胖的男性DM患者应警惕CRC发生;CRC发生风险与DM病程呈正相关;DM会增加CRC发生风险,且其可使CRC患者病情恶化及淋巴结或血行转移风险增加。     

EGFR 19-Del上调Fn14及JAK/STAT的表达促进非小细胞肺癌的发生发展

目的:本研究检测Fn14、p-JAK1、p-STAT1在EGFR 19-Del的非小细胞肺癌(non-small cell lung cancer,NSCLC)细胞系中的表达,并探讨EG F R 1 9-Del对Fn 1 4及J A K 1/STAT 1表达的调控作用,以期为EGFR 19-Del之NSCLC的发展机制研究奠定基础。方法:采用EGFR TKI(吉非替尼)处理HCC827细胞系(EGFR 19-Del)、H1975(L858R)及H292细胞系(正常肺上皮细胞),Western blot法检测Fn14、p-JAK1、p-STAT1蛋白表达。结果:相比于H1975和H292细胞系,Fn14、p-JAK1及p-STAT1基因和蛋白在HCC827细胞系中有较高的表达水平。对比未经吉非替尼处理的HCC827细胞系,经抑制剂处理后的HCC827细胞系中Fn14、p-JAK1、p-STAT1蛋白表达明显受到抑制,而在H292细胞系中无此现象。由此说明,EGFR 19-Del对Fn14及JAK/STAT信号分子的表达具有调控作用。结论:EGFR 19-Del可能通过上调Fn14及JAK/STAT信号分子的表达促进NSCLC的发生发展,而Fn14可能是EGFR 19-Del之NSCLC潜在的治疗靶点。     

微小RNA在结直肠癌中的研究进展

结直肠癌(colorectal carcinoma,CRC)是消化系统最常见的恶性肿瘤之一,随着中国生活条件和饮食习惯的改变,CRC发病率呈明显上升趋势。因此,对CRC发生、发展的机制以及早期诊断标志物的研究是目前CRC研究的重点。自研究者在秀丽隐杆线虫(C.elegans)中发现了第1个定时调控胚胎后期     

成纤维细胞生长因子诱导14在结直肠中的表达及其临床意义

目的:探讨成纤维细胞生长因子诱导14(Fn14)在结直肠癌中的表达及其临床意义。方法:应用免疫组化SP法检测Fn14在结直肠正常黏膜上皮细胞、腺瘤及腺癌中的表达状况。结果:Fn14在结直肠腺癌中表达率为56.7%,显著高于散发性腺瘤(7.1%)及正常结直肠黏膜上皮细胞的表达率(0%),差异有显著性(P<0.01)。Fn14蛋白表达与淋巴结转移相关(P<0.05),与病理组织分期及年龄、性别无相关性(P>0.05)。结论:Fn14的异常表达与结直肠腺癌的发生及淋巴结转移相关。     

叉头框蛋白M1过表达在结直肠癌中的作用

目的:评估叉头框蛋白M1(forkhead box protein M1,FOXM1)在结直肠癌(colorectal cancer,CRC)的发生发展中的作用。方法:研究103例原发CRC和配对的正常组织标本,探讨FOXM1表达变化的潜在机制及其对体外CRC细胞模型的增殖和转移的影响。结果:103例CRC组织染色后FOXM1阳性率为85.44%(88/103),癌旁正常组织中阳性率为20.39%(21/103)。两组间的FOXM1的表达差异具有统计学意义(P0.001);FOXM1沉默能抑制结肠癌细胞的增殖,且侵袭和迁移也明显受抑。结论:CRC的发病机制可能是由FOXM1介导,FOXM1可能代表CRC分子靶向治疗的选择性靶点。     

甲状腺激素T3影响K562细胞转铁蛋白受体和铁蛋白表达的分子机制研究

目的 探讨甲状腺激素T3对白血病细胞K562转铁蛋白受体(TfR)和铁蛋白(Fn)表达的调控作用及其可能的机制。方法 采用流式细胞术和放射免疫法分别检测TfR和Fn的表达水平，用RNA/蛋白带移动分析法测定铁调节蛋白(IRP)与铁反应元件(IRE)的结合活性。应用RT—PCR方法半定量分析TfR和Fn的mRNA水平。结果 T3对K562细胞TfR的表达无明显作用，而以不同浓度的T3处理细胞后使Fn的表达增加，其中当T3为100nmol/L和200nmol/L时，与空白对照组相比，差异有显性(P＜0．05)。T3能减少IRP/IRE的结合活性，且以T3为50nmol/L时减少最为明显。不同浓度的T3在不同的作用时间内均能提高H—FnmRNA水平，而对TfRmRNA水平无显影响。结论 T3可增加细胞Fn的表达，而对TfR的表达无明显调控作用，其机制除包含转录后的调节外，可能还具有转录水平的调控作用。     

Panax notoginseng saponins radiosensitize colorectal cancer cells by regulating the SNHG6/miR-137 axis

Panax notoginseng saponins (PNS) have recently attracted great attention for their anti-cancer activity in colorectal cancer (CRC). The aim of this study was to explore the functional role and underlying mechanisms of PNS on CRC radiosensitivity. Cell viability was assessed by a Cell Counting kit-8 assay. Cell survival and apoptosis were determined using colony formation assay and flow cytometry, respectively. Quantitative real-time PCR was used to quantify the levels of SNHG6 and miR-137. The targeted correlation between SNHG6 and miR-137 was validated by dual-luciferase reporter and RNA immunoprecipitation assays. Our data supported that PNS weakened the viability of CRC cells. Moreover, PNS promoted the radiosensitivity of CRC cells. Mechanistically, PNS enhanced CRC cell radiosensitivity by upregulating SNHG6. SNHG6 directly targeted miR-137 and inhibited miR-137 expression. MiR-137 was involved in the regulatory effect of SNHG6 on CRC cell radiosensitivity. Furthermore, PNS increased miR-137 expression through SNHG6 in CRC cells. Our study suggested that PNS promoted radiosensitivity in CRC cells at least partly through regulating the SNHG6/miR-137 axis, providing a novel understanding of the anti-cancer mechanism of PNS in CRC.

Panax notoginseng saponins (PNS) have recently attracted great attention for their anti-cancer activity in colorectal cancer (CRC).     

Clinical impact of Fn-induced high expression of KIR2DL1 in CD8 T lymphocytes in oesophageal squamous cell carcinoma

BackgroundTo analyze the correlation between the inducing effect of Fusobacterium nucleatum (Fn) on the surface expression of the inhibitory receptor KIR2DL1 on CD8⁺ T cells in oesophageal squamous cell carcinoma (ESCC) and the clinicopathological features and survival prognosis and to explore its clinical significance.MethodsThe inducing effect of Fn on CD8⁺ T cell surface inhibitory receptor KIR2DL1 expression was analyzed in a coculture system of human CD8⁺ T cells and ESCC cells infected with Fn. Fn infection and the expression of KIR2DL1 on CD8⁺ T cells were detected by RNAscope and immunohistochemistry in ESCC tissues, and the correlations between the inducing effect of Fn on KIR2DL1 expression on CD8⁺ T cells and clinicopathological features were analyzed. COX regression was used to analyze the influence of each factor on the prognosis of ESCC. Survival curves were plotted by the Kaplan–Meier method, and the effect of KIR2DL1 induction on survival time was analyzed by the log-rank test.ResultsIn the coculture system, KIR2DL1 expression on the surface of CD8⁺ T cells increased with increasing Fn infection time. In ESCC tissues, Fn infection was significantly correlated with high KIR2DL1 expression on CD8⁺ T cells. The Fn + CD8⁺KIR2DL1 positive patients were predominantly males who were smokers and alcohol drinkers. Moreover, patients with Fn infection were characterized by poor tumour differentiation, advanced clinical stage, and a short survival time. Meanwhile, Fn + CD8⁺KIR2DL1 positive group was independent risk factor affecting the prognosis of ESCC patients.ConclusionsLong-term drinking and smoking lead to an extremely unhealthy oral environment in which Fn infection and colonization are more likely to occur, thus inducing high expression of KIR2DL1 on the surface of CD8⁺ T cells, which can weaken the antitumour immune response and promote the malignant progression of ESCC.

HIGHLIGHTS

• Fn induced high expression of KIR2DL1 CD8⁺ T cells in a time-dependent manner.
• Fn can reduce the response of tumour cells to CDDP.
• The inducing effect of Fn on CD8⁺ T cell surface KIR2DL1 expression was significantly associated with the poor prognosis of ESCC patients.

     

miR-142-3p靶向Wnt/β-链蛋白通路对结直肠癌细胞增殖的影响

目的探究miR-142-3p靶向Wnt/β-链蛋白(β-catenin)通路对结直肠癌(CRC)细胞增殖的影响。方法选取2018年1月—2020年10月收治的66例CRC的肿瘤组织及其相邻正常组织。同时培养正常人结肠上皮细胞NCM460和CRC细胞系(HT29、LoVo、HCT116、Caco2、SW480细胞)。通过qRT-PCR检测CRC肿瘤组织、正常组织、正常人结肠上皮细胞与CRC细胞系中miR-142-3p表达水平;经免疫蛋白印迹法检测CRC细胞系中β-catenin表达水平;采用CCK-8法和流式细胞术检测miR-142-3p过表达后对CRC细胞增殖的影响;经免疫蛋白印迹检测过表达miR-142-3p对Wnt/β-catenin信号通路相关蛋白的影响;采用双荧光素酶报告基因检验miR-142-3p与β-catenin编码基因CTNNB1的靶向关系。结果miR-142-3p在CRC肿瘤组织和CRC细胞系中的表达显著下降(P<0.05);β-catenin在CRC细胞系中的表达显著升高(P<0.05);过表达miR-142-3p可靶向结合CTNNB1,显著抑制Caco2、LoVo和HT29细胞的增殖和Ki67+细胞比例,抑制β-catenin、c-myc和Cyclin D1的表达(P<0.05)。结论miR-142-3p可通过靶向调节β-catenin的表达,干扰Wnt/β-catenin通路,抑制CRC细胞的增殖。     

Circular RNA hsa_circ_0000567 can be used as a promising diagnostic biomarker for human colorectal cancer

Background

Recent studies have revealed that circular RNAs are involved in the biological process of some kinds of human cancers. However, little is known about their diagnostic values and functions in colorectal cancer (CRC).

Methods

The expression levels of hsa_circ_0000567 in 102 paired CRC tissues and adjacent noncancerous tissues, 5 CRC cell lines, and a normal colorectal epithelial cell line were detected by quantitative real‐time polymerase chain reaction (qRT‐PCR). The correlations between hsa_circ_0000567 expression levels and the clinicopathological factors of patients with CRC were analyzed. Furthermore, the loss‐of‐function assay was performed to investigate the functions of hsa_circ_0000567 in vitro. Finally, a receiver operating characteristic (ROC) curve was established to evaluate the diagnostic value of hsa_circ_0000567.

Results

Hsa_circ_0000567 expression was significantly downregulated in CRC tissues and CRC cell lines. In addition, the decreased hsa_circ_0000567 expression in CRC was negatively correlated with tumor size (P = .011), lymph metastasis (P = .003), distal metastasis (P < .0001), and tumor–node–metastasis (TNM) stage (P = .003) in CRC. Moreover, knockdown of hsa_circ_0000567 promoted CRC cells proliferation and migration in vitro. Importantly, the area under the ROC curve (AUC) was 0.8653, which indicates hsa_circ_0000567 can serve as a diagnostic biomarker.

Conclusion

Hsa_circ_0000567 may be a novel suppressor and a potential diagnosis biomarker in CRC.

     

Development and characterization of a potent immunoconjugate targeting the Fn14 receptor on solid tumor cells

TNF-like weak inducer of apoptosis (TWEAK) and fibroblast growth factor (FGF)-inducible 14 (Fn14) are a TNF superfamily ligand-receptor pair involved in many cellular processes including proliferation, migration, differentiation, inflammation, and angiogenesis. The Fn14 receptor is expressed at relatively low levels in normal tissues, but it is known to be dramatically elevated in a number of tumor types, including brain and breast tumors. Thus, it seems to be an excellent candidate for therapeutic intervention. We first analyzed Fn14 expression in human tumor cell lines. Fn14 was expressed in a variety of lines including breast, brain, bladder, skin, lung, ovarian, pancreatic, colon, prostate, and cervical cancer cell lines. We then developed an immunoconjugate containing a high-affinity anti-Fn14 monoclonal antibody (ITEM-4) conjugated to recombinant gelonin (rGel), a highly cytotoxic ribosome-inactivating N-glycosidase. Both ITEM-4 and the conjugate were found to bind to cells to an equivalent extent. Confocal microscopic analysis showed that ITEM4-rGel specifically and rapidly (within 2 hours) internalized into Fn14-positive T-24 bladder cancer cells but not into Fn14-deficient mouse embryonic fibroblasts. Cytotoxicity studies against 22 different tumor cell lines showed that ITEM4-rGel was highly cytotoxic to Fn14-expressing cells and was 8- to 8 × 10(4)-fold more potent than free rGel. ITEM4-rGel was found to kill cells by inducing apoptosis with high-mobility group box 1 protein release. Finally, ITEM4-rGel immunoconjugate administration promoted long-term tumor growth suppression in nude mice bearing T-24 human bladder cancer cell xenografts. Our data support the use of an antibody-drug conjugate approach to selectively target and inhibit the growth of Fn14-expressing tumors.     

KITENIN-targeting MicroRNA-124 Suppresses Colorectal Cancer Cell Motility and Tumorigenesis

MicroRNAs are increasingly implicated in the modulation of the progression of various cancers. We previously observed that KAI1 C-terminal interacting tetraspanin (KITENIN) is highly expressed in sporadic human colorectal cancer (CRC) tissues and hence the functional KITENIN complex acts to promote progression of CRC. However, it remains unknown that microRNAs target KITENIN and whether KITENIN-targeting microRNAs modulate CRC cell motility and colorectal tumorigenesis. Here, through bioinformatic analyses and functional studies, we showed that miR-124, miR-27a, and miR-30b negatively regulate KITENIN expression and suppress the migration and invasion of several CRC cell lines via modulation of KITENIN expression. Through in vitro and in vivo induction of mature microRNAs using a tetracycline-inducible system, miR-124 was found to effectively inhibit the invasion of CT-26 colon adenocarcinoma cells and tumor growth in a syngeneic mouse xenograft model. Constitutive overexpression of precursor miR-124 in CT-26 cells suppressed in vivo tumorigenicity and resulted in decreased expression of KITENIN as well as that of MYH9 and SOX9, which are targets of miR-124. Thus, our findings identify that KITENIN-targeting miR-124, miR-27a, and miR-30b function as endogenous inhibitors of CRC cell motility and demonstrate that miR-124 among KITENIN-targeting microRNAs plays a suppressor role in colorectal tumorigenesis.     

Role of bile acids in colon carcinogenesis

Bile acids (BAs) are cholesterol derivatives synthesized in the liver and then secreted into the intestine for lipid absorption. There are numerous scientific reports describing BAs, especially secondary BAs, as strong carcinogens or promoters of colon cancers. Firstly, BAs act as strong stimulators of colorectal cancer (CRC) initiation by damaging colonic epithelial cells, and inducing reactive oxygen species production, genomic destabilization, apoptosis resistance, and cancer stem cells-like formation. Consequently, BAs promote CRC progression via multiple mechanisms, including inhibiting apoptosis, enhancing cancer cell proliferation, invasion, and angiogenesis. There are diverse signals involved in the carcinogenesis mechanism of BAs, with a major role of epidermal growth factor receptor, and its down-stream signaling, involving mitogen-activated protein kinase, phosphoinositide 3-kinase/Akt, and nuclear factor kappa-light-chain-enhancer of activated B cells. BAs regulate numerous genes including the human leukocyte antigen class I gene, p53, matrix metalloprotease, urokinase plasminogen activator receptor, Cyclin D1, cyclooxygenase-2, interleukin-8, and miRNAs of CRC cells, leading to CRC promotion. These evidence suggests that targeting BAs is an efficacious strategies for CRC prevention and treatment.     

Studies on the fibronectin receptors of human peripheral blood leukocytes. Morphologic and functional characterization

We have investigated the interactions between plasma fibronectin (Fn) and human peripheral blood phagocytic cells. As shown by studies of the binding of Fn-coated fluorescent microspheres (Fn-ms), both polymorphonuclear leukocytes (PMN) and monocytes had specific binding sites for Fn at the plasma membrane. However, as purified from blood, only monocytes were stimulated by Fn to become more actively phagocytic. This increase in phagocytosis was reflected by an Fn- induced increase in the ingestion of IgG-coated erythrocytes and, more dramatically by an Fn-dependent initiation of phagocytosis of C3b- coated erythrocytes. Despite this difference between PMN and monocytes in the functional consequences of Fn binding, the cell surface molecules responsible for Fn binding on the two cell types shared many characteristics. On both cells, binding of Fn-ms was inhibited by sufficient concentrations of fluid-phase Fn; both PMN and monocytes bound fewer Fn-ms at 4 degrees C than at 37 degrees C; both achieved maximal binding at similar Fn-ms/cell ratios; and phenylmethylsulfonyl fluoride did not inhibit Fn-ms binding to either cell type. Most dramatically, monoclonal anti-Fn antibodies that inhibited binding of Fn-ms to one cell type inhibited binding to both; conversely, monoclonal anti-Fn antibodies that did not inhibit Fn-ms binding to either cell type did not inhibit binding to the other. Fn will stimulate PMN to a more actively phagocytic state, like that induced in monocytes, if the PMN are first exposed to C5a or N-formyl-methionyl- leucylphenylalanine. This effect occurs without apparent change in the number of Fn receptors. We conclude that the PMN and monocyte receptors for Fn are very similar, but that their milieu is very different in the two cells as purified from peripheral blood. Whereas Fn induces increased phagocytosis in monocytes, PMN must be activated before the Fn can be effective.