The microRNA-520a-3p inhibits proliferation,apoptosis and metastasis by targeting MAP3K2 in non-small cell lung cancer

Growing evidence indicates that miR-520a was involved in the complement attack and migration of tumor cells, but nonetheless, the role of miR-520a-3p in non-small cell lung cancer (NSCLC) is not clear. Mitogen-activated protein kinase kinase kinase 2 (MAP3K2) is a kinase belonging to the serine/threonine protein kinase family. To develop potential therapy targeting MAP3K2, we studied the roles of miR-520a-3p in the proliferation, apoptosis and metastasis of NSCLC. The expression levels of miR-520a-3p were quantified in tumor tissues of NSCLC by qRT-PCR, and the mimics and inhibitors were used to verify the function of miR-520a-3p. The cell proliferation was evaluated by MTT assay, and the migration and invasion was evaluated by transwell assay. The athymic mice subcutaneous injection was used to research NSCLC cell tumor formation. The bioinformatics tools and luciferase assay was applied to detect the relationship between miR-520a-3p and its target. Protein levels of miR-520a-3p target was determined by western blot analysis. MiR-520a-3p expression was decreased in the NSCLC tissues compared with their normal counterparts and lower expression of miR-520a-3p in NSCLC tissues was associated with a higher clinical stage, NSCLC metastasis and poor prognosis. Inhibition of expression of miR-520a-3p can reduce in vitro NSCLC cell migration and invasion as well as in vivo metastasis. MAP3K2 mRNA contains a binding site for miR-520a-3p in the 3’UTR. MAP3K2 is one of target of miR-520a-3p. Together, our data demonstrated that miR-520a-3p inhibits proliferation, apoptosis and metastasis in NSCLC by targeting MAP3K2, and miR-520a-3p may be used as a prognosis marker for NSCLC in clinical research.     

卵巢上皮恶性肿瘤侵袭转移相关miRNA的筛选与鉴定

目的 探索与卵巢上皮恶性肿瘤侵袭转移可能相关的miRNA.方法 采用miRNA芯片,筛选SKOV-3ip和SKOV-3细胞差异表达miRNA.利用生物信息学软件TargetScan、MicroCosm、PicTar和GO,预测差异表达miRNA的靶基因及其功能.采用实时逆转录聚合酶链反应(real-time RT-PCR)技术,验证与卵巢癌侵袭转移可能相关的5种miRNA(let-7a、let-7e、let-7f、miR-22和miR-886-5p)在SKOV-3ip和SKOV-3细胞的表达.同时,检测这5种miRNA在另外一组侵袭转移能力不同的卵巢癌细胞株HO8910和HO-8910PM中的表达,并进行统计学分析.结果 基因芯片筛选显示,42种miRNA在SKOV-3ip和SKOV-3细胞株表达差异明显.进一步分析显示,let-7a、let-7e、let-7f、miR-22和miR-886-5p等5种miRNA可能与卵巢癌的侵袭转移密切相关.real-time RT-PCR结果证实,let-7f和miR-22在两组侵袭转移能力不同的卵巢癌细胞(SKOV-3和SKOV-3ip细胞、HO-8910和HO-8910PM细胞)表达差异有统计学意义(均P＜0.05).结论 let-7f和miR-22在侵袭转移能力强的卵巢癌细胞中低表达,可能具有抑癌基因的作用.     

SMYD3 promotes implant metastasis of ovarian cancer via H3K4 trimethylation of integrin promoters

Detachment of cancer cells from the primary tumor and formation of spheroids in ascites is required for implantation metastasis in epithelial ovarian cancer (EOC), but the underlying mechanism of this process has not been thoroughly elucidated. To mimic this process, ovarian cancer cells were grown in 3D and 2D culture. Hey and OVCA433 spheroids exhibited decreased cell proliferation and enhanced adhesion and invasion. SMYD3 expression was elevated in ovarian carcinoma spheroids in association with increased H3K4 methylation. Depletion of SMYD3 by transient siRNA, stable shRNA knockdown and the SMYD3 inhibitor BCI-121 all decreased spheroid invasion and adhesion. Gene expression arrays revealed downregulation of integrin family members. Inhibition assays confirmed that invasion and adhesion of spheroids are mediated by ITGB6 and ITGAM. SMYD3-deficient cells regained the ability to invade and adhere after forced overexpression of SMYD3, ITGB6 and ITGAM. However, this biological ability was not restored by forced overexpression of SMYD3 in ITGB6- and/or ITGAM-deficient cancer cells. SMYD3 and H3K4me3 binding at the ITGB6 and ITGAM promoters was increased in spheroids compared to that in monolayer cells, and the binding was decreased when SMYD3 expression was inhibited, consistent with the expression changes in integrins. SMYD3 expression and integrin-mediated adhesion were also activated in an intraperitoneal xenograft model and in EOC patient spheroids. In vivo, SMYD3 knockdown inhibited tumor metastasis and reduced ascites volume in both the intraperitoneal xenograft model and a PDX model. Overall, our results suggest that the SMYD3-H3K4me3-integrin pathway plays a crucial role in ovarian cancer metastasis to the peritoneal surface.     

Differential expression of CXCR4 is associated with the metastatic potential of human non-small cell lung cancer cells.

PURPOSE: To evaluate the relation between CXCR4 expression and the presence of metastatic disease in human non-small cell lung cancer (NSCLC) patients and investigate whether modulation of CXCR4 expression could serve as a potential pathway in preventing metastasis of NSCLC. EXPERIMENTAL DESIGN: CXCR4 expression in 36 patients with NSCLC and 10 normal lung tissues was detected by real-time PCR and immunohistochemistry. CXCR4 expression in two human NSCLC clones (95C and 95D) with different metastatic potential was determined by real-time PCR and flow cytometry. 95C and 95D cells were transfected with the plasmid DNA containing CXCR4 coding gene or CXCR4 antisense nucleotide fragment, respectively, and the effects on in vitro cell migration, invasion, and adhesion and in vivo metastasis were measured. RESULTS: Up-regulated expression of CXCR4 was detected in 34 tumors, which were further divided into 17 high expression cancers and 17 low expression cancers by their staining intensities. High CXCR4 tumors (13 of 17) were more prone to clinical metastasis in comparison with low expression tumors. CXCR4 was differentially expressed in 95C and 95D cells with low or high metastatic potential, and the surface expression of CXCR4 were 50% up-regulated or down-regulated following the stable transfection. The metastatic potential of NSCLC in vitro, such as migration, invasion, and adhesion, were significantly enhanced or impaired. In addition, neutralizing the interactions of stromal cell-derived factor-1/CXCR4 in vitro with CXCR4-specific antibodies inhibited the CXCR4-dependent migration, invasion, and adhesion. Furthermore, s.c. inoculation of lung cancer cells with low expression of CXCR4 in nude mice showed 0- to 2-fold decrease in lung metastatic foci than that with high expression of CXCR4. CONCLUSIONS: Differential expression of CXCR4 is associated with the metastatic potential of human NSCLC, raising the possibility that blockade of CXCR4/stromal cell-derived factor-1 interaction may lead the way to design novel therapeutic tools for the treatment of metastatic NSCLC patients.     

Interfering with ITGB1-DT expression delays cancer progression and promotes cell sensitivity of NSCLC to cisplatin by inhibiting the MAPK/ERK pathway

Long non-coding RNA ITGB1-DT is involved in the regulation of cancer growth and metastasis. However, the roles of ITGB1-DT in non-small cell lung cancer (NSCLC) progression and sensitivity to cisplatin has not been elucidated. ITGB1-DT expression in NSCLC tissues, and the relationship between ITGB1-DT expression with NSCLC diagnosis, prognosis, clinicopathological features, and immune cell infiltration were investigated in The Cancer Gene Atlas (TCGA) database. The roles and mechanisms of ITGB1-DT in cell growth, migration, and drug sensitivity of NSCLC cells were explored in the cell model. The prognostic nomograms of ITGB1-DT-related genes were evaluated using bioinformatics. ITGB1-DT was overexpressed in NSCLC. Elevated ITGB1-DT expression was related to the late T stage, N stage, M stage, short overall survival (OS), disease-specific survival (DSS), and progression-free interval (PFI) of NSCLC patients. ITGB1-DT was the independent risk factors for poor prognosis, and had diagnostic value for NSCLC patients. Interfering with the ITGB1-DT expression can inhibit the proliferation, migration, and invasion of A549, H1299, and drug-resistant A549/DDP, possibly due to the inhibition of p38 MAPK and ERK phosphorylation levels. ITGB1-DT expression was correlated with the levels of NSCLC immune infiltration cells, such as the TReg, Th, and NK cells. ITGB1-DT-related gene nomograms were associated with the prognosis, and were expected to evaluate the prognosis of NSCLC patients. In conclusion, inhibition of ITGB1-DT expression delayed the growth and metastasis of NSCLC using the MAPK/ERK signaling mechanism and enhanced the sensitivity of NSCLC to cisplatin drugs. These results indicate that ITGB1-DT might be a biomarker for evaluating the diagnosis and prognosis of NSCLC patients.     

MiR-449a suppresses cell invasion by inhibiting MAP2K1 in non-small cell lung cancer

Increasing evidence reveals that deregulation of miRNAs contributes to carcinogenesis of the human non-small cell lung cancer (NSCLC). Our study discovered that the expression of miR-449a was markedly decreased in NSCLC cells with high metastatic capacity and tissues of positive lymph node metastasis. Moreover, our results showed that miR-449a could act as a tumor suppressor by inhibiting the invasion of NSCLC cells in vitro and in vivo. Mechanistically, miR-449a inhibited the expression of MAP2K1 by direct targeting its 3’UTR, and regulated the activity of MEK1/ERK1/2/c-Jun pathway through an auto-regulatory feedback loop. Furthermore, the histone methylation mediated the decreased expression of miR-449a through SUZ12. Taken together, the novel connection between miR-449a and MAP2K1 demonstrated here provided a new, potential therapeutic target for the treatment of non-small cell lung cancer.     

Let-7c Inhibits NSCLC Cell Proliferation by Targeting HOXA1

Objective: The aim of the present study was to explore mechanisms by which let-7c suppresses NSCLC cellproliferation. Methods: The expression level of let-7c was quantified by qRT-PCR. A549 and H1299 cells weretransfected with let-7c mimics to restore the expression of let-7c. The effects of let-7c were then assessed by cellproliferation, colony formation and cell cycle assay. Mouse experiments were used to confirm the effect of let-7con tumorigenicity in vivo. Luciferase reporter assays and Western blotting were performed to identify targetgenes for let-7c. Results: HOXA1 was identified as a novel target of let-7c. MTS, colony formation and flowcytometry assays demonstrated that forced expression of let-7c inhibited NSCLC cell proliferation by inducingG1 arrest in vitro, consistent with inhibitory effects induced by knockdown of HOXA1. Mouse experimentsdemonstrated that let-7c expression suppressed tumorigenesis. Furthermore, we found that let-7c could regulatethe expression of HOXA1 downstream effectors CCND1, CDC25A and CDK2. Conclusions: Collectively, theseresults demonstrate let-7c inhibits NSCLC cell proliferation and tumorigenesis by partial direct targeting ofthe HOXA1 pathway, which suggests that restoration of let-7c expression may thus offer a potential therapeuticintervention strategy for NSCLC.     

miRNA-30b调控ITGB3表达促进乳腺癌细胞凋亡

  目的  探讨微小RNA-30b （microRNA-30b,miRNA-30b）和整合素β3（integrin β3,ITGB3）在乳腺癌组织中的表达及对乳腺癌细胞生物学行为的影响。  方法  选择2016年3月至2019年3月于成都医学院第一附属医院收治的144例包括原位、浸润性和转移性乳腺癌患者的组织标本以及其癌旁组织,同时收集其临床病理资料。采用实时荧光定量PCR（RT-PCR）检测miRNA-30b和ITGB3的表达。构建miRNA-30b高表达的miRNA-30b模拟物和ITGB3高表达的miRNA-30b模拟物-pc DNA3.1-ITGB3,转染乳腺癌MCF-7细胞,同时将转染后的细胞皮下注射至小鼠体内。5-溴脱氧尿嘧啶核苷（BrdU）实验、流式细胞术、Transwell小室侵袭实验检测细胞增殖、凋亡、侵袭,Western blot检测细胞中的ITGB3表达。  结果  RT-PCR结果显示,原位、浸润性和转移性乳腺癌组织中的miRNA-30b相对表达量分别为0.75、0.52和0.23,ITGB3 mRNA分别为2.17、3.76和5.43,与正常组织相比,差异均具有统计学意义（均P<0.001）。miRNA-30b高表达后,BrdU、流式细胞术、Transwell小室侵袭实验、Western blot检测结果显示,癌细胞的增殖、侵袭能力以及ITGB3表达明显降低,凋亡率升高（均P<0.001）。  结论  在乳腺癌组织中miRNA-30b和ITGB3表达异常,miRNA-30b过表达能明显下调ITGB3表达,抑制乳腺癌细胞的增殖、侵袭,并促使其凋亡。      

慢病毒介导β4整合素shRNA对人非小细胞肺癌H460SM细胞皮下移植瘤生长的影响

目的:研究慢病毒介导的β4整合素(integrinβ4,ITGB4) shRNA对人非小细胞肺癌(non-small cell lung cancer,NSCLC)裸鼠皮下移植瘤生长的抑制作用.方法:采用实时荧光定量-PCR (real-time fluorogenic quantitative-PCR,RFQ-PCR)和蛋白质印迹法检测ITGB4在不同人NSCLC细胞中的表达水平;慢病毒介导ITGB4 shRNA抑制NSCLC H460SM细胞中ITGB4的表达,再用RFQ-PCR和蛋白质印迹法检测ITGB4的表达水平,评价基因沉默效率;裸鼠随机分为3组,皮下分别接种H460SM-NS细胞(对照组)、稳定表达ITGB4 shRNA的H460SM-68和H460SM-71细胞.每隔1d测量裸鼠体质量和肿瘤大小,比较各组裸鼠肿瘤生长情况.处死裸鼠,剥离肿瘤,测量肿瘤大小和质量.结果:ITGB4在大多数人NSCLC细胞中均有表达,其中大细胞肺癌H460SM和NCI-H460细胞中ITGB4表达量明显高于NCI-H661细胞(P＜0.01),H460SM细胞中ITGB4表达水平明显高于亲本NCI-H460细胞(P＜0.01):ITGB4 shRNA转染组细胞中ITGB4的表达水平明显低于阴性对照组细胞(P＜0.01); ITGB4 shRNA转染组皮下移植瘤生长速度明显低于阴性对照组(P＜0.01).结论:ITGB4表达可能与人NSCLC细胞的致瘤、侵袭、转移的表型有关;抑制ITGB4的表达,可以抑制人NSCLC细胞H460SM裸鼠皮下移植瘤的生长.     

NOX4 promotes non-small cell lung cancer cell proliferation and metastasis through positive feedback regulation of PI3K/Akt signaling

NADPH oxidase 4 (NOX4) is deregulated in various cancers and involved in cancer proliferation and metastasis. However, what the role of NOX4 plays during malignant progression of non-small cell lung cancer (NSCLC) remains unknown. Our results show that NOX4 was upregulated in NSCLC cell lines and samples from patients, compared with controls; NOX4 protein levels were closely correlated with clinical disease stage and survival time. Overexpression of NOX4 in A549 and H460 NSCLC cells enhanced cell proliferation and invasion in vitro, and produced larger tumors, shorter survival time, and more lung metastasis in nude mice than control cells. On the contrary, NOX4 depletion inhibited NSCLC cell aggressiveness. Inhibition of PI3K/Akt pathway could sufficiently block the cellular effects of NOX4 overexpression in NSCLC cells both in vitro and in vivo. Specifically, we demonstrated that PI3K/Akt pathway also positively regulated NOX4 expression via NF-κB-mediated manner. Therefore, there existed a mutual positive regulation between NOX4 and PI3K/Akt signaling in NSCLC cells, and NOX4 was confirmed to functionally interplay with PI3K/Akt signaling to promote NSCLC cell proliferation and invasion. In conclusion, the positive feedback loop between NOX4 and PI3K/Akt signaling contributes to NSCLC progression.     

The regulatory mechanism of CCR7 gene expression and its involvement in the metastasis and progression of gastric cancer

Gastric cancer is the second leading cause of cancer mortality, but the molecular mechanisms underlying its progression and metastasis remain unclear. CCR7 and Dicer 1 protein expression in 80 gastric adenocarcinomas and 40 peritumoral tissues were measured by immunohistochemical staining. The expression of let-7a miRNA in serum, tumor tissues, and peritumoral tissues was measured by real-time PCR. The role of let-7a in CCR7 protein expression, migration, and invasion of gastric cancer cells was tested in vitro. Dicer 1 protein expression was found to be significantly reduced, whereas CCR7 protein expression was significantly increased in gastric adenocarcinomas compared to peritumoral tissues. The let-7a miRNA levels in the serum and tumor tissues of gastric adenocarcinoma patients were significantly lower than in the serum of healthy controls and peritumoral tissues, respectively. Dicer 1 protein positively correlated with let-7a miRNA level, but negatively correlated with CCR7 protein level in gastric adenocarcinoma. Negative Dicer 1 protein and let-7a miRNA expression and positive CCR7 protein expression significantly correlated with lymph node metastasis, depth of invasion, high clinical TNM stage, and larger tumor size. Let-7a transfection significantly inhibited CCR7 protein expression, migration, and invasion of MNK-45 cells in vitro. High expression of CCR7 protein and low expression of Dicer 1 protein and let-7a miRNA are significantly associated with the metastasis and progression of gastric cancer. High CCR7 protein expression may be caused by the loss of Dicer 1 protein expression and reduced let-7a miRNA level in gastric cancer. The serum let-7a level might be a marker for the diagnosis of gastric cancer.     

Pivotal role of reduced let-7g expression in breast cancer invasion and metastasis

Screening of the entire let-7 family of microRNAs (miRNA) by in situ hybridization identified let-7g as the only member, the diminished expression of which was significantly associated with lymph node metastasis and poor survival in breast cancer patients. Abrogation of let-7g expression in otherwise nonmetastatic mammary carcinoma cells elicited rapid metastasis from the orthotopic location, through preferential targets, Grb2-associated binding protein 2 (GAB2) and fibronectin 1 (FN1), and consequent activation of p44/42 mitogen-activated protein kinase (MAPK) and specific matrix metalloproteinases. Treatment with estrogen or epidermal growth factor specifically reduced the expression of mature let-7g through activation of p44/42 MAPK and subsequently stimulated expression of GAB2 and FN1, which, in turn, promoted tumor invasion. We thus identify let-7g as a unique member of the let-7 miRNA family that can serve as a prognostic biomarker in breast cancer and also propose a paradigm used by specific signaling molecules via let-7g to cooperatively promote breast cancer invasion and metastasis. Thus, let-7 family members neither possess equivalent clinicopathologic correlation nor function in breast cancer.     

Reduced sialidase expression in highly metastatic variants of mouse colon adenocarcinoma 26 and retardation of their metastatic ability by sialidase overexpression.

Sialidase expression levels are inversely correlated with the metastatic potential of mouse colon adenocarcinoma 26 sublines, as assessed by activity assays and RT-PCR, irrespective of total and cell surface sialic acid contents. Compared with low metastatic NL4 and NL44 cell lines, the highly metastatic NL17 and NL22 cells exhibit low expression of sialidases, accompanied with higher levels of sialylLe(x) and GM3. To investigate whether these properties of NL17 cells can be altered by sialidase overexpression, we transfected a cytosolic sialidase gene into NL17 cells. The result was markedly inhibited lung metastasis, invasion and cell motility with a concomitant decrease in sialylLe(x) and GM3 levels, in line with the case of spontaneously low metastatic sublines having relatively high endogenous sialidase levels, implying that sialidase level is a determining factor affecting metastatic ability. Treatment of the cells with antibodies against sialylLe(x) and GM3 affected cell adhesion and/or cell motility, providing evidence that desialylation of these molecules, as targets of sialidase, is involved in the suppression of metastasis.     

PTEN/PI3K信号转导相关蛋白在非小细胞肺癌组织中的表达及其意义

背景与目的:PTEN/PI3K信号转导途径能调节细胞的增殖与存活,与多种肿瘤的发生发展密切相关,但在肺癌中所起的作用及其相互关系尚不十分明确。本研究旨在探讨PTEN/PI3K信号转导相关蛋白IGF-1R、PTEN和PI3K在非小细胞肺癌(non-smallcelllungcarcinoma,NSCLC)中的表达及其意义。方法:利用免疫组织化学SP法检测59例NSCLC组织、19例转移的淋巴结组织、16例癌旁增生肺组织和7例正常肺组织中IGF-1R、PTEN和PI3K蛋白的表达。IGF-1R、PTEN和PI3K蛋白阳性率的差异比较采用卡方检验和Fisher确切概率法。结果:59例NSCLC组织中IGF-1R、PTEN和PI3K蛋白阳性率分别为:72.88%、27.12%和84.75%。IGF-1R以及PI3K蛋白的阳性率与NSCLC的临床病理特征无显著相关(P>0.05)。PTEN蛋白的阳性率与NSCLC的组织学类型、细胞分化程度无关(P>0.05),但与淋巴结转移显著相关(P=0.009)。NSCLC组及转移淋巴结组中PI3K、PTEN蛋白的阳性率与正常肺组织和癌旁增生肺组织相比,差异均有显著性(P<0.05)。59例NSCLC组织中IGF和PI3K之间具有显著正相关性(P=0.001),Pearson列联系数为0.432。PTEN和PI3K之间具有显著负相关(P=0.000),Pearson列联系数为0.505。结论:IGF-1R、PI3K蛋白的过表达和PTEN蛋白的低表达与肺癌的发生及浸润转移相关。     

趋化因子CXC受体3在肝细胞癌中的表达及意义

背景与目的:最近研究揭示趋化因子及其受体网络在肿瘤侵袭转移中起重要作用。本研究旨在探讨趋化因子CXC受体3(CXCR3)在肝细胞癌(hepatocellularcarcinoma,HCC)中的表达,及其与HCC临床病理特征的关系。方法:选取7株人HCC细胞系、18例正常肝组织、64例HCC患者的癌组织和癌旁肝组织作为研究对象。采用RT-PCR和实时定量PCR方法,研究这些组织中CXCR3mRNA的表达情况。采用免疫组织化学方法,研究CXCR3蛋白的表达情况和HCC复发转移的关系。结果:在高转移细胞系MHCC97-H中,CXCR3与!2微球蛋白的mRNA拷贝均数之比为33.0×10-4,在低转移细胞系MHCC97-L中,该数值为8.7×10-4,在无转移能力的5株细胞系则未检出CXCR3的mRNA表达。在MHCC97-H和MHCC97-L中均见CXCR3蛋白强阳性染色,而在无转移能力的细胞系中只有HepG2见到微弱染色,其余4株均未见染色。CXCR3蛋白在HCC组织中的强阳性染色,与HCC的侵袭性正相关(P=0.003)。HCC组织中CXCR3蛋白强阳性的患者和CXCR3阴性或弱阳性的患者的1、2年无瘤生存率分别为66.7%、31.3%和75.0%、59.5%,差异有显著性(P=0.044)。结论:CXCR3的表达与HCC的侵袭和转移相关。     

Snail和Claudin-3在非小细胞肺癌中的表达及意义

背景与目的 上皮-间质转化( epithelial mesenchymal transition,EMT)是肿瘤浸润和转移的关键步骤,上皮细胞极性丧失是其主要标志,表现为Claudin等上皮标记丢失.锌指转录因子Snail是调控EMT的重要转录因子,近年来对肿瘤侵袭转移机制研究发现Snail能提高多种肿瘤的侵袭能力.本研究旨在利用组织芯片技术探讨转录因子Snail和紧密连接蛋白(Claudin-3)在非小细胞肺癌(non-small cell lung cancer,NSCLC)及其淋巴结转移灶中的表达和意义.方法 分别采用免疫组织化学MaxVision法和EnVision法检测59例癌旁正常肺组织、302例NSCLC原发灶以及57例淋巴结转移灶中Snail和Claudin-3的表达.结果 Snail在癌旁正常肺组织、NSCLC原发灶以及淋巴结转移灶中的表达逐渐增强,差异具有统计学意义(P＜0.05);Claudin-3在癌旁正常肺组织、NSCLC原发灶以及淋巴结转移灶中的表达逐渐减弱,差异具有统计学意义(P＜0.05).在NSCLC原发灶中,Snail和Claudin-3的表达与肿瘤组织学类型有关(P＜0.05).Spearman等级相关分析显示Snail与Claudin-3的表达呈负相关(r=-0.178,P=0.002).Kaplan-Meier生存分析显示肿瘤大小、组织学类型、病理分级、有无癌转移、TNM分期、Snail的表达以及Snail与Claudin-3的差异性表达影响NSCLC患者的术后生存时间(P＜0.05).Cox回归分析提示肿瘤大小、组织学类型、病理分级、有无癌转移和TNM分期是影响NSCLC患者预后的独立危险因素(P＜0.05).结论 Snail和Claudin-3在NSCLC的浸润、转移中具有重要意义,有助于对NSCLC患者预后的评价.     

血管内皮钙黏蛋白对非小细胞肺癌侵袭转移的影响及其作用机制

目的 探讨非小细胞肺癌（non-small cell lung cancer, NSCLC）组织中血管内皮钙黏蛋白（vascular endothelial cadherin, VE-cadherin）的表达和其对细胞侵袭迁移能力的影响及作用机制。方法 采用免疫组织化学、RT-PCR法检测VE-cadherin在人NSCLC组织和癌旁组织中的表达;Westernblot法检测VE-cadherin在NSCLC细胞株中的表达;Transwell实验检测VE-cadherin对NCI-H460细胞侵袭迁移能力的影响;免疫荧光检测VE-cadherin与P120ctn在NCI-H460细胞上的定位;免疫共沉淀检测VE-cadherin与P120ctn的连接情况。结果 VE-cadherin在肺癌组织中表达水平较癌旁组织上调,其表达与NSCLC有无吸烟史、有无淋巴结转移有关（P<0.05）。VE-cadherin在PG、NCI-H460、 A549细胞株表达水平不同,干扰VE-cadherin表达后,NCI-H460细胞侵袭迁移能力下调,低氧可诱导VEcadherin在NCI-H460细胞表达增强,VE-cadherin与P120ctn存在结构上的连接。结论 VE-cadherin可能通过缺氧信号活化及P120ctn介导参与NSCLC侵袭、转移。     

MicroRNA let-7a represses chemoresistance and tumourigenicity in head and neck cancer via stem-like properties ablation

Head and neck cancer (HNC) is a prevalent cancer worldwide. Let-7 has been shown to function as a tumour suppressor by regulating multiple oncogenic signalling pathways. However, the role of let-7 in head and neck cancer (HNC) and in HNC-associated tumour initiating cells (TIC) remains unclear. In this study, we first demonstrated that let-7a expression was significantly decreased but that Nanog/Oct4 expression was increased in HNC tissues as compared to adjacent normal cells. Expression of let-7a in recurrent HNC tissue and in regional metastatic lymph nodes of HNC patients was also significantly decreased, but Nanog/Oct4 expression was increased as compared to the expression levels in the parental tumours. Consistently, the stemness genes were significantly up-regulated and let-7a was down-regulated in HNC-ALDH1(+) cells relative to HNC-ALDH1(-) cells. Furthermore, lentiviral-mediated let-7a overexpression could significantly inhibit the stemness signature and the chemoresistant abilities of HNC-ALDH1(+) cells. Most importantly, overexpression of let-7 or knockdown of Nanog in ALDH1(+) cells effectively blocked tumour metastasis and significantly prolonged survival time in ALDH1(+)-transplanted immunocompromised mice. Overall, restoration of let-7a in HNC and HNC-TIC may be a new approach for the therapeutic treatment of HNC in the future. These results show that let-7a negatively modulates the expression of stemness genes and plays a role as a tumour suppressor in HNC by eliminating the putative HNC-TIC population.     

MTA家族蛋白表达与非小细胞肺癌侵袭转移的相关性探讨

目的:探讨肿瘤转移相关基因(matastasis associated gene,MTA)家族蛋白作为非小细胞肺癌（non-small cell lung cancer,NSCLC）预后预测因子的价值,为进一步了解该基因家族与肿瘤侵袭和转移相关的分子机制提供参考。方法:应用免疫组化(SABC)检测MTA家族（MTA1、MTA2、MTA3）蛋白在54例NSCLC肿瘤组织中的表达水平,分析与NSCLC临床病理特征的关系;选取正常肺上皮细胞BEAS-2B、肺癌高转移细胞株95-D与低转移细胞株95-C,对比MTA1在其中的表达与细胞侵袭转移的相关性。结果:MTA1、MTA2和MTA3在NSCLC组织中均有较高的表达,而MTA1表达水平与淋巴结转移状态相关;细胞学实验结果显示MTA1在肿瘤细胞中高表达,而MTA1在肺癌高转移细胞95-D中的表达水平显著高于低转移细胞95-C。结论:MTA1表达水平与肿瘤侵袭、转移关系密切,可考虑作为肿瘤转移、预后预测因子。