GJA1在结直肠癌组织中表达及促进侵袭转移的研究

摘要 目的：间隙连接Alpha-1蛋白（Gap Junction Alpha-1,GJA1）是间隙连接中分布最广泛的蛋白,并在多种肿瘤中起促癌作用,但其在结直肠癌发生、发展的作用研究甚少。本实验旨在探究GJA1在结直肠癌组织中的表达情况及其对结直肠癌细胞系侵袭、转移能力的影响,以期为结直肠癌的诊断和预后寻找新的生物标志物。方法：收集92对结直肠癌及其癌旁组织样本,提取组织RNA,利用qRT-PCR检测GJA1相对表达量,并分析GJA1表达与临床病理特征及预后的相关性。在HCT116和HCT8两种结直肠癌细胞系中分别构建GJA1过表达载体和敲减载体,利用qRT-PCR和、Western Blot检测上皮间充质转化（epithelial-mesenchymal transition, EMT）相关蛋白E-Cadherin、N-Cadherin、Vimentin和Snail的表达变化,利用Wound healing和Transwell实验观察其迁移、侵袭能力的变化。结果：相对于癌旁组织,GJA1在结直肠癌组织中显著低表达。并且结直肠癌中低表达的GJA1与肿瘤分化程度、浸润深度、淋巴血管转移相关,低表达GJA1结直肠癌患者显示更差的总体生存率和更低的无病生存率。此外,过表达GJA1后,结直肠癌细胞E-cadherin的表达升高,N-cadherin、Vimentin和Snail的表达降低,划痕愈合减慢,Transwell转移细胞减少;而敲减GJA1后,结直肠癌细胞E-Cadherin的表达降低,N-Cadherin、Vimentin和Snail的表达升高,划痕愈合加快,Transwell转移细胞增多。结论：GJA1在结直肠癌中低表达,其表达降低可通过EMT促进结直肠癌的侵袭、转移并影响病人预后。     

SATB1 Expression Is Associated with Biologic Behavior in Colorectal Carcinoma In Vitro and In Vivo

There is increasing evidence that Special AT-rich sequence-binding protein 1 (SATB1) is aberrantly expressed in several cancers and is correlated with clinicopathologic parameters in these tumors. In this study, we showed over-expression of SATB1 in 80 cases of colorectal cancer and in 3 colorectal cancer cell lines and found expression levels were strongly associated with tumor differentiation and stage. Expression levels of SATB1 protein were higher in poorly-differentiated as compared with well-differentiated cell lines, and both quantity and distribution patterns of SATB1 were associated with tumor differentiation and pTNM stage. Strikingly, we further investigated the effect of down regulation of SATB1 expression on malignant phenotypic features in colorectal cancer cells in vitro, and showed that SABT1 down-regulation negatively affected growth potential, anchorage-independent colony formation and cancer cell invasion, and resulted in increased apoptosis. SATB1 expression was positively associated with the expression of various biological and genetic markers, including Cyclin D1, MMP-2, NF-κB, and PCNA, and was associated with loss of APC and BRAF^V600E. These findings suggest that SATB1 is involved in the carcinogenesis, development and progression of colorectal cancer.     

IgG Expression in Human Colorectal Cancer and Its Relationship to Cancer Cell Behaviors

Increasing evidence indicates that various cancer cell types are capable of producing IgG. The exact function of cancer-derived IgG has, however, not been elucidated. Here we demonstrated the expression of IgG genes with V(D)J recombination in 80 cases of colorectal cancers, 4 colon cancer cell lines and a tumor bearing immune deficient mouse model. IgG expression was associated with tumor differentiation, pTNM stage, lymph node involvement and inflammatory infiltration and positively correlated with the expressions of Cyclin D1, NF-κB and PCNA. Furthermore, we investigated the effect of cancer-derived IgG on the malignant behaviors of colorectal cancer cells and showed that blockage of IgG resulted in increased apoptosis and negatively affected the potential for anchor-independent colony formation and cancer cell invasion. These findings suggest that IgG synthesized by colorectal cancer cells is involved in the development and growth of colorectal cancer and blockage of IgG may be a potential therapy in treating this cancer.     

丝甘蛋白聚糖促进非小细胞肺癌的侵袭与转移

丝甘蛋白聚糖(serglycin,SRGN)在肿瘤细胞的侵袭与转移中具有广泛的研究前景。本研究报道SRGN与肺癌细胞侵袭与转移能力之间的相关性。首先,通过检测SRGN在正常肺上皮细胞株BEAS-2B及不同侵袭与转移能力的肺癌细胞株95C、95D中的表达差异。利用shRNA干扰技术,在侵袭与转移能力强的95D细胞中建立稳定干扰SRGN表达的95D/shSRGN的细胞株,并通过RT-qPCR、Western印迹、酶联免疫吸附测定验证其敲除效率。结果显示：干扰SRGN可抑制侵袭与转移性强的95D细胞的侵袭与转移能力,减弱细胞迁移与侵袭等生物学特性,导致上皮标志物上皮细胞钙黏连蛋白（E-cadherin）表达上调,间质标志物纤维连接蛋白1（fibronectin1, FN1）及EMT（epithelial-mesenchymal transition, EMT）相关转录因子锌指E盒结合同源框1（zinc finger E-box binding homeobox 1, ZEB1）表达下调。进一步分析发现, SRGN与上皮细胞钙黏连蛋白表达成负相关（P=-0.25）,而与FN1(P=0.12)及ZEB1(P=0.35)表达成正相关,并且SRGN高表达的患者总生存时间明显少于SRGN低表达组（P=0.0077）,SRGN与ZEB1同时高表达的患者,总生存时间显著小于SRGN与ZEB1低表达患者（P=0.0005）。研究结果证实,SRGN促进上皮间质转化发生,增强非小细胞肺癌的侵袭与转移能力,为非小细胞肺癌预后提供参考。     

B7-H1 Expression Is Associated with Poor Prognosis in Colorectal Carcinoma and Regulates the Proliferation and Invasion of HCT116 Colorectal Cancer Cells

Background And Objective

The investigation concerning the B7-H1 expression in colorectal cancer cells is at an early stage. It is unclear whether B7-H1 expression may have diagnostic or prognostic value in colorectal carcinoma. Additionally, how B7-H1 is associated with the clinical features of colorectal carcinoma is not known. In order to investigate the relationship between B7-H1 and colorectal cancer, we analyzed B7-H1 expression and its effect in clinical specimens and HCT116 cells.

Methods

Paraffin-embedded specimens from 143 eligible patients were used to investigate the expression of CD274 by immunohistochemistry. We also examined whether B7-H1 itself may be related to cell proliferation, apoptosis, migration and invasion in colon cancer HCT116 cells.

Results

Our results show that B7-H1 was highly expressed in colorectal carcinoma and was significantly associated with cell differentiation status and TNM (Tumor Node Metastasis) stage. Patients with positive B7-H1 expression showed a trend of shorter survival time. Using multivariate analysis, we demonstrate that positive B7-H1 expression is an independent predictor of colorectal carcinoma prognosis. Our results indicate that B7-H1 silencing with siRNA inhibits cell proliferation, migration and invasion. Furthermore, cell apoptosis was also increased by B7-H1 inhibition.

Conclusions

Positive B7-H1 expression is an independent predictor for colorectal carcinoma prognosis. Moreover, knockdown of B7-H1 can inhibit cell proliferation, migration and invasion.     

Cyclin D1 regulates lung cancer invasion and metastasis

Cyclin D1, as a regulatory factor in cell cycle, is highly expressed in many tumors, such as lung cancer, breast cancer and thyroid cancer. The aim of the present study was to study the role of Cyclin D1 in invasion and metastasis of lung cancer cells. Lung adenocarcinoma cell line A549 and squamous cell line SK-MES-1 were selected as the objects, because A549 expresses Cyclin D1 highly, and SK-MES-1 expresses lowly. Nude mice were injected with A549 or SK-MES-1 via tail vein, and were sacrificed after 4 weeks for cancer tissue isolation. The harvested cancer cells were reinjected into another nude mouse. After one more time of such seeding, highly metastatic lung cancer model was established. After A549 and SK-MES-1 were transfected with Cyclin D1 RNAi and expression vector respectively, transwell migration assay was used to analyze transferring capacity of lung cancer cells. Western blot was used to detect Cyclin D1 and WNT/TCF pathway proteins expressions in parental cell lines and cancer tissue from metastasis model animals. The results showed that, along with the increase of seeding times, lung cancer cells from model animals, no matter A549 or SK-MES-1, exhibited augmented metastasis activity and up-regulated Cyclin D1 expression. The transferring capacity was weakened significantly in A549 cells where the Cyclin D1 was interfered by RNAi, and it was enhanced significantly in SK-MES-1 cells which were transfected with the expression vector of Cyclin D1. The expressions of WNT/TCF pathway proteins, including β-catenin, lymphoid enhancer-binding factor (LEF) and T cell factor (TCF), increased significantly in highly metastatic model animals. The parental cell lines showed lower expressions of WNT/TCF pathway proteins compared with cancer tissue from metastasis model animals. These results suggest that Cyclin D1 is closely related with the invasion and metastasis of lung cancer cells, and the WNT/TCF signal pathway may promote the expression of Cyclin D1.     

CCNB2在结直肠癌中的表达及临床意义

目的:探讨细胞周期蛋白B2(Cyclin B2,CCNB2)在结直肠癌组织中的表达及其临床意义。方法:选择45对结直肠癌组织及癌旁正常结直肠组织样本,分别采用实时定量PCR(qRT-PCR)方法和免疫组织化学技术检测CCNB2的mRNA和蛋白表达,并进一步分析CCNB2的表达与结直肠癌临床病理特征之间的关系。结果:结直肠癌组织中CCNB2 mRNA的表达显著高于癌旁正常结直肠组织,差异有统计学意义(P0.001),且CCNB2的mRNA表达与结直肠癌的肿瘤大小、浸润深度及TNM分期显著相关(P0.05),与年龄、性别、肿瘤位置、分化程度、脉管神经浸润、淋巴结转移和远处转移均无关(P0.05)。45例结直肠癌标本中39例表达(+~+++),6例表达(-)。CCNB2蛋白主要表达于结直肠癌细胞质中,少量见于细胞核。结直肠癌组织中CCNB2蛋白的阳性表达率为86.7%,显著高于癌旁正常结直肠组织,并与患者的性别、年龄、分化程度和肿瘤转移均无显著相关性(P0.05),但与肿瘤分期、浸润程度均显著相关(P0.05)。结论:CCNB2在结直肠癌中呈异常高表达,且与结直肠癌的发生发展相关,有望作为结直肠癌的诊断和预后预测参考指标。     

L1-mediated colon cancer cell metastasis does not require changes in EMT and cancer stem cell markers

Aberrant activation of Wnt/β-catenin signaling is common in most sporadic and inherited colorectal cancer (CRC) cells leading to elevated β-catenin/TCF transactivation. We previously identified the neural cell adhesion molecule L1 as a target gene of β-catenin/TCF in CRC cells. Forced expression of L1 confers increased cell motility, invasion, and tumorigenesis, and the induction of human CRC cell metastasis to the liver. In human CRC tissue, L1 is exclusively localized at the invasive front of such tumors in a subpopulation of cells displaying nuclear β-catenin. We determined whether L1 expression confers metastatic capacities by inducing an epithelial to mesenchymal transition (EMT) and whether L1 cosegregates with cancer stem cell (CSC) markers. We found that changes in L1 levels do not affect the organization or expression of E-cadherin in cell lines, or in invading CRC tissue cells, and no changes in other epithelial or mesenchymal markers were detected after L1 transfection. The introduction of major EMT regulators (Slug and Twist) into CRC cell lines reduced the levels of E-cadherin and induced fibronectin and vimentin, but unlike L1, Slug and Twist expression was insufficient for conferring metastasis. In CRC cells L1 did not specifically cosegregate with CSC markers including CD133, CD44, and EpCAM. L1-mediated metastasis required NF-κB signaling in cells harboring either high or low levels of endogenous E-cadherin. The results suggest that L1-mediated metastasis of CRC cells does not require changes in EMT and CSC markers and operates by activating NF-κβ signaling.     

Down-regulation of MUC1 in cancer cells inhibits cell migration by promoting E-cadherin/catenin complex formation

MUC1, a tumor associated glycoprotein, is over-expressed in most cancers and can promote proliferation and metastasis. The objective of this research was to study the role of MUC1 in cancer metastasis and its potential mechanism. Pancreatic (PANC1) and breast (MCF-7) cancer cells with stable 'knockdown' of MUC1 expression were created using RNA interference. beta-Catenin and E-cadherin protein expression were upregulated in PANC1 and MCF-7 cells with decreased MUC1 expression. Downregulation of MUC1 expression also induced beta-catenin relocation from the nucleus to the cytoplasm, increased E-cadherin/beta-catenin complex formation and E-cadherin membrane localization in PANC1 cells. PANC1 cells with 'knockdown' MUC1 expression had decreased in vitro cell invasion. This study suggested that MUC1 may affect cancer cell migration by increasing E-cadherin/beta-catenin complex formation and restoring E-cadherin membrane localization.     

TFCP2L1在子宫内膜癌细胞增殖及侵袭中的作用

摘要 目的：研究人子宫内膜癌中TFCP2L1的表达情况以及分析TFCP2L1对子宫内膜癌细胞增殖及迁移能力的影响。方法：（1）通过TCGA 及GTEx数据库分析子宫内膜癌中TFCP2L1的表达水平及患者生存期。采用Western blot验证正常子宫内膜上皮细胞与多种子宫内膜癌细胞系中TFCP2L1的表达情况。（2）使用CRISPR-Cas9技术敲除Ishikawa细胞系的TFCP2L1,并用流式分选技术筛选单个细胞进行培养,形成单克隆细胞系,以此来研究TFCP2L1对子宫内膜癌的细胞周期和细胞增殖的影响。通过 Western blot 及细胞免疫荧光检测细胞周期相关蛋白的表达,检测细胞增殖情况,采用平板克隆实验及CCK8实验。（3）通过Transwell小室及划痕实验对侵袭和转移能力进行检测。结果：TCGA 及GTEx数据库分析发现TFCP2L1在子宫内膜癌中高表达且与肿瘤患者预后不良相关。敲除TFCP2L1后,Ki67、Cyclin D1 和 Cyclin D2的蛋白水平显著下调,CCK8及平板克隆实验结果表明,敲除TFCP2L1能够显著降低子宫内膜癌细胞的增殖能力。划痕实验及Transwell侵袭实验结果表明敲除TFCP2L1的子宫内膜癌细胞侵袭迁移能力均减弱。结论：本研究证明了TFCP21L1是子宫内膜癌的促癌因子。TFCP2L1的高表达可能与子宫内膜癌预后不良相关。敲除TFCP2L1可以抑制子宫内膜癌的侵袭和转移。     

N-myc Downstream Regulated Gene 1 (NDRG1) Promotes Metastasis of Human Scirrhous Gastric Cancer Cells through Epithelial Mesenchymal Transition

Our recent study demonstrated that higher expression of N-myc downregulated gene 1 (NDRG1) is closely correlated with poor prognosis in gastric cancer patients. In this study, we asked whether NDRG1 has pivotal roles in malignant progression including metastasis of gastric cancer cells. By gene expression microarray analysis expression of NDRG1 showed the higher increase among a total of 3691 up-regulated genes in a highly metastatic gastric cancer cell line (58As1) than their parental low metastatic counterpart (HSC-58). The highly metastatic cell lines showed decreased expression of E-cadherin, together with enhanced expression of vimentin and Snail. This decreased expression of E-cadherin was restored by Snail knockdown in highly metastatic cell lines. We next established stable NDRG1 knockdown cell lines (As1/Sic50 and As1/Sic54) from the highly metastatic cell line, and both of these cell lines showed enhanced expression of E-cadherin and decreased expression of vimentin and Snail. And also, E-cadherin promoter-driven luciferase activity was found to be increased by NDRG1 knockdown in the highly metastatic cell line. NDRG1 knockdown in gastric cancer cell showed suppressed invasion of cancer cells into surround tissues, suppressed metastasis to the peritoneum and decreased ascites accumulation in mice with significantly improved survival rates. This is the first study to demonstrate that NDRG1 plays its pivotal role in the malignant progression of gastric cancer through epithelial mesenchymal transition.     

Silencing of lncRNA ZFAS1 inhibits malignancies by blocking Wnt/β-catenin signaling in gastric cancer cells

Gastric cancer is a common malignancy with high mortality. Long noncoding RNA (lncRNA) zinc finger antisense (ZFAS)1 is upregulated in gastric cancer specimens compared with the para-carcinoma tissues. The silencing of ZFAS1 inhibited the growth, proliferation, cell cycle progress, migration, invasion and epithelial-mesenchymal transition (EMT), and enhanced the sensitivity to cis-platinum or paclitaxel in SGC7901 cells, as evidenced by the expression changes of proliferating cell nuclear antigen, Cyclin D1, Cyclin E, Cyclin B1, E-cadherin, N-cadherin, vimentin, matrix metalloproteinase (MMP)-2 and MMP-14. The ZFAS1 also activated the Wnt/β-catenin signaling. Subsequently, the ZFAS1 knockdown-induced the inhibition of migration, invasion, EMT and resistance to chemotherapeutic reagens was reversed by the overexpression of β-catenin. In summary, the silencing of ZFAS1 inhibited the growth, proliferation, cell cycle progress, migration, invasion, EMT and chemotherapeutic tolerance by blocking the Wnt/β-catenin signaling in gastric cancer cells.     

Matrix metalloproteinase 2 promotes cell growth and invasion in colorectal cancer

Colorectal cancer (CRC) is the third leading cause of cancer-related death in the western world. In this study, we evaluated the expression of matrix metalloproteinase 2 gene (MMP2) in CRC and analyzed its correlation with clinicopathological features. We found that the expression of MMP2 was significantly higher in CRC tissues than in the colorectal tissues. In addition, high levels of MMP2 protein were positively correlated with the status of tumor size, lymph node metastasis, distant metastasis, Dukes' stage, and tumor invasion. Moreover, patients with higher MMP2 levels had markedly shorter overall survivals than those with low MMP2 levels. Multivariate analysis results suggested that the level of MMP2 expression is an independent prognostic indicator for the survival of patients with CRC. Silencing MMP2 expression in CRC cell lines with lentiviral-mediated shRNA markedly suppressed cell proliferation, colony formation, and invasion. Furthermore, we observed that vascular endothelial growth factor (VEGF) and membrane type 1 (MT1)-MMP protein levels were decreased in MMP2-down-regulated colorectal cells. Therefore, our study demonstrated that MMP2 is an important factor related to carcinogenesis and metastasis of CRC, and MMP2 promotes CRC cell growth and invasion by up-regulating VEGF and MT1-MMP expression, which makes this pathway a potential target for cancer treatment.     

SALL4 as an Epithelial-Mesenchymal Transition and Drug Resistance Inducer through the Regulation of c-Myc in Endometrial Cancer

SALL4 plays important roles in the development and progression of many cancers. However, the role and molecular mechanism of SALL4 in endometrial cancer remain elusive. In the present research, we have demonstrated that the expression of SALL4 was upregulated in endometrial cancer and correlated positively with tumor stage, metastases and poor survival of patients. The overexpression of SALL4 promoted the invasiveness in endometrial cancer cells, as indicated by the upregulation of mesenchymal cell marker N-cadherin and downregulation of the epithelial marker E-cadherin, and invasion assays in vitro. Additionally, there was also an increase in drug resistance in these cell models due to the upregulation of ATP-binding cassette multidrug transporter ABCB1 expression. Moreover, we also found that ABCB1 was critical for SALL4-induced drug resistance. In contrast, SALL4 knockdown restored drug sensitivity, reversed EMT, diminished cell metastasis and suppressed the downregulation of E-cadherin and the upregulation of N-cadherin and ABCB1. Furthermore, we showed that SALL4 upregulated c-Myc expression and c-Myc was a direct target for SALL4 by ChIP assay, depletion of c-Myc with siRNA abolished the SALL4-induced downregulation of E-cadherin, upregulation of N-cadherin and ABCB1, suggesting that c-Myc was a downstream target for SALL4 and required for SALL4-induced EMT, invasion and drugs resistance in endometrial cancer cells. These results indicated that SALL4 could induce EMT and resistance to antineoplastic drugs through the regulation of c-Myc. SALL4 and c-Myc may be novel therapeutic targets for endometrial cancer.     

ZC3H13 suppresses colorectal cancer proliferation and invasion via inactivating Ras–ERK signaling

ZC3H13 is a canonical CCCH zinc finger protein, which harbors a somatic frame-shift mutation in colorectal cancer (CRC). However, its expression and biological function were still uncertain. In the current study, we found that ZC3H13 was served as a tumor suppressor in CRC cells, which decreased the expression of Snail, Cyclin D1, and Cyclin E1, and increased the expression of Occludin and Zo-1 through inactivating Ras–ERK signaling pathway. Furthermore, reduction of ZC3H13 associated with advanced TNM stage (p = 0.02), positive regional lymph node metastasis ( p = 0.01). Taken together, the current study indicated that ZC3H13 may be an upstream regulator of Ras–ERK signaling pathway and suppressed invasion and proliferation of CRC.     

FMNL2 enhances invasion of colorectal carcinoma by inducing epithelial-mesenchymal transition

FMNL2 is a member of diaphanous-related formins that control actin-dependent processes such as cell motility and invasion. Its overexpression in metastatic cell lines and tissues of colorectal carcinoma has been associated with aggressive tumor development in our previous study. But its specific role in cancer is largely unknown. Here we report that FMNL2 is involved in epithelial-mesenchymal transition (EMT) maintenance in human colorectal carcinoma cells. A positive correlation between FMNL2 and vimentin expression and an inverse correlation between FMNL2 and E-cadherin expression were found in colorectal carcinoma cell lines and cancer tissues. Specific knockdown of FMNL2 led to an epithelial-state transition, confirmed by the cobblestone-like phenotype, upregulation of E-cadherin, α-catenin, and γ-catenin, and downregulation of vimentin, snail, slug. Loss of FMNL2 expression lowered the ability of TGF-β to induce cell invasion and EMT, as shown by morphology and the expression levels. Upregulation of vimentin, slug, snail, downregulation of E-cadherin and activation of receptor-Smad3 phosphorylation were observed in M5 and MDCK cells induced by TGF-β, whereas altered expression of these markers was not obvious in FMNL2-depleting M5 cells. High levels of activation of p-MAPK and p-MEK, but not p-PI3K and p-AKT, were observed in SW480/FMNL2+ cells compared with control cells. Treatment with U0126 could abrogate the activation of p-MAPK and p-MEK, whereas LY294002 treatment had no effect on the PI3K/AKT pathway. In conclusion, these findings identify a novel EMT and tumor promoting function for FMNL2, which is involved in TGF-β-induced EMT and colorectal carcinoma cell invasion via Smad3 effectors, or in collaboration with MAPK/MEK pathway.     

AEG-1对大肠癌进展的影响

目的研究AEG-1在大肠癌组织和细胞中的表达,探讨AEG-1通过调控上皮间质转化和耐药参与大肠癌的进展。方法采用qRT-PCR检测AEG-1在大肠癌组织和细胞中的表达,统计AEG-1对大肠癌患者生存率的影响,分析其在不同癌症分期患者中的表达差异,并分析AEG-1表达量与大肠癌诊断敏感性的关系。采用体外实验将si-NC、pc-DNA-NC、si-AEG-1、pc-DNA-AEG-1转染到大肠癌SW116和LOVO细胞中,然后通过qRT-PCR检测转染效率以及AEG-1在两细胞系中的表达情况。采用CCK-8和克隆形成实验检测AEG-1对大肠癌细胞增殖的影响;采用流式细胞术检测转染后上皮间质转化和耐药情况的变化;采用Western blotting检测转染后上皮间质转化和耐药相关蛋白N-cadherin、E-cadherin、MRP的变化情况。结果 77例大肠癌患者组织中AEG-1表达水平明显高于对照组。浸润T3+T4期的患者中AEG-1的表达水平高于浸润T1+T2期患者。Ⅲ+Ⅳ期患者中AEG-1的表达水平高于Ⅰ+Ⅱ期。AEG-1高表达组患者OS时间明显低于低表达组。AEG-1表达量与大肠癌诊断敏感性之间呈显著正相关。在LOVO细胞系中,降低AEG-1表达后其细胞活力、侵袭力明显降低,同时间质细胞标志蛋白N-cadherin、上皮细胞标志蛋白E-cadherin、多药耐药相关蛋白MRP表达量降低。在SW116细胞系中,过表达AEG-1后上皮间质转化、耐药相关蛋白表达量显著升高。结论 AEG-1在大肠癌组织和细胞中的表达量明显上升,AEG-1通过调控上皮间质转化和耐药参与大肠癌的发生发展,为大肠癌的治疗提供了新的理论依据和新的靶点。     

沉默FOXQ1可逆转SW480细胞的上皮-间质转化

FOXQ1是FOX家族的的重要成员之一,其参与了多种人类肿瘤的上皮间质转化(epithelial- mesenchymal transition,EMT).本研究设计合成了FOXQ1基因的shRNA（short hairpin RNA）,用此转染SW480细胞,通过显微镜观察细胞形态,Transwell小室、细胞黏附试验检测转移能力及黏附能力,以探索FOXQ1与结直肠癌细胞EMT的关系.结果显示,沉默FOXQ1后,SW480细胞顶底极性及细胞间紧密连接增加,侵袭、迁移的细胞数目减少,同种黏附能力增加,异种黏附能力降低.进一步的机制研究表明,沉默FOXQ1基因可以导致SW480细胞的上皮标志因子E-cadherin表达显著增高,而间质细胞标志因子N-cadherin、Vimentin及MMP2表达均降低.以上结果表明,沉默FOXQ1基因可以逆转SW480细胞EMT,其机制可能与E-cadherin的上调和N cadherin、Vimentin、MMP2的下调有关,这为进一步研究FOXQ1在结直肠癌发生发展中的作用提供实验基础.