Troglitazone induces G1 arrest by p27(Kip1) induction that is mediated by inhibition of proteasome in human gastric cancer cells.

We examined in the present study whether human gastric cancer cells express peroxisome proliferator-activated receptor gamma (PPARgamma), the effect of PPARgamma activation by troglitazone, a selective ligand, on cellular growth, and the mechanism of the growth arrest by troglitazone in gastric cancer cells. RT-PCR, northern blot and western blot analysis demonstrated that all four tested human gastric cancer cell lines, MKN-28, MKN-45, MKN-74 and KATO-III, expressed PPARgamma mRNA and protein. WST-1 assay and flow cytometric analysis revealed that troglitazone inhibited the growth and induced G1 arrest in all four gastric cancer cell lines. To examine the role of p27(Kip1), a cyclin-dependent kinase inhibitor, in the G1 arrest by troglitazone, we determined p27(Kip1) protein expression by western blot analysis in gastric cancer cells that had been treated with troglitazone. Troglitazone increased p27(Kip1) in all four gastric cancer cell lines. Since it has been reported that the ubiquitin-proteasome system plays a vital role in the degradation of p27(Kip1) protein, we evaluated the hypothesis that inhibition of proteasome mediates the troglitazone-induced p27(Kip1) accumulation. Lactacystin, a proteasome inhibitor, inhibited cell growth and increased p27(Kip1) expression in MKN-74 cells. It was further demonstrated that troglitazone inhibited proteasome activity in a dose-dependent manner in MKN-74 cells. All these results suggest that troglitazone inhibited proteasome activity, followed by induction of p27(Kip1), which arrests cells at the G1 phase of the cell cycle in gastric cancer cells. The troglitazone-mediated inhibition of the proteasome suggests a novel mechanism for the anti-proliferative effect of this agent in cancer cells.     

MEMO1共结合IGF-IR介导胃癌细胞侵袭转移

目的:探讨MEMO1（mediator of ErbB2-driven cell motility 1）调控IGF-IR（insulin-like growth factor-I receptor）介导胃癌侵袭转移的分子机制,为临床治疗提供潜在靶点。方法:选取五种不同分化程度胃癌细胞系（MKN-7、NCI-N87、MGC-803、BGC-823、MKN-74）,Western blot检测MEMO1、IGF-IR和E-cad表达差异;Transwell比较五种胃癌细胞系迁移能力;选取MEMO1/IGF-IR高表达细胞系MGC-803,瞬时敲除MEMO1或IGF-IR基因,Western blot检测EMT标志物表达差异;IP实验检测MEMO1与IGF-IR间相互作用;TCGA数据库分析胃癌组织MEMO1和IGF-IR mRNA表达水平与患者预后的关系。结果:基础状态下五种胃癌细胞IGF-IR与MEMO1的表达呈显著正相关,并且两种蛋白的表达强度与胃癌细胞EMT相关;五种胃癌细胞迁移能力MKN-7相似文献   

Effects of the Hippo Signaling Pathway in Human Gastric Cancer

Background/Aim: The Hippo signaling pathway is a newly discovered and conserved signaling cascade,which regulates organ size control by governing cell proliferation and apoptosis. This study aimed to investigateits effects in human gastric cancer. Methods: Tumor tissues (n=60), adjacent non-tumor tissues (n=60) andnormal tissues (n=60) were obtained from the same patients with primary gastric cancer (GC). In addition,70 samples of chronic atrophic gastritis (CAG) tissues were obtained from patients with intestinal metaplasia(IM) by endoscopic biopsy. Hippo signaling molecules, including Mst1, Lats1, YAP1, TAZ, TEAD1, Oct4 andCDX2, were determined by quantitative polymerase chain reaction (qPCR). Protein expression of Mst1, Lats1,YAP1, TEAD1 and CDX2 was assessed by immunohistochemistry and Western blotting. Results: Mst1, Lats1and Oct4 mRNA expression showed an increasing tendency from GC tissues to normal gastric tissues, while themRNA expression of YAP1, TAZ and TEAD1 was up-regulated (all P<0.01). Mst1 and Lats1 protein expressionpresented a similar trend with their mRNA expression. In addition, YAP1 and TEAD1 protein expression in GCwas significantly higher than in the other groups (all P<0.01). CDX2 mRNA and protein expression in the CAGgroup were higher than in the other groups (all P<0.01). In GC, mRNA expression of Mst1, Lats1, Oct4, YAP1,TAZ, TEAD1 and CDX2 had a close correlation with lymphatic metastasis and tumor TNM stage (all P<0.01).Furthermore, protein expression of Mst1, Lats1 ,YAP1, TAZ, TEAD1 and CDX2 had a close correlation betweeneach other (P<0.05). Conclusion: The Hippo signaling pathway is involved in the development, progression andmetastasis of human gastric cancer. Therefore, manipulation of Hippo signaling molecules may be a potentialtherapeutic strategy for gastric cancer.     

Docetaxel enhances the cytotoxicity of cisplatin to gastric cancer cells by modification of intracellular platinum metabolism

We have examined the combined anticancer effects of docetaxel (DOC) and cisplatin (CDDP) in vitro using the gastric cancer cell lines MKN-45, MKN-74, and TMK-1. Treatment of the cell lines with 30 microg/ml of DOC for 24 h followed by incubation with 3 or 10 microg/ml of CDDP for 24 h showed a clear synergistic effect. Sequence dependency of the agents was observed in these cell lines: DOC followed by CDDP (DC) showed a stronger antitumor effect than CDDP followed by DOC (CD) in all cell lines. To clarify the mechanism of action of the DC combination, total intracellular platinum (Pt) levels were evaluated after treatment with CDDP alone or combined with DC. For the MKN-45 and -74 cell lines, cells treated with DOC (10 microg/ml for 12 h) and then CDDP showed significantly increased intracellular Pt accumulation compared to cells treated with CDDP alone. We also investigated alterations in intracellular glutathione (GSH) concentration in response to DOC and CDDP. MKN-45 and -74 cells pretreated with DOC (10 microg/ml for 12 h) showed significantly increased intracellular GSH levels compared to cells administered CDDP only. To explain these findings, messenger RNA (mRNA) levels for multidrug resistance-associated protein-1 (MRP-1), the ATP-dependent pump for Pt-GSH complexes, were quantified in CDDP-treated MKN-45 cells with and without DOC pretreatment. While CDDP administration increased MRP-1 mRNA expression in MKN-45 cells, MRP-1 was not up-regulated after CDDP administration in DOC pretreated MKN-45 cells. Our results suggested that the enhanced CDDP toxicity due to DOC pretreatment may be related to the accumulation of intracellular Pt-GSH complexes, because DOC appears to suppress the MRP-1 up-regulation induced by CDDP exposure in gastric cancer cells.     

Genetic Status and Expression of the Cyclin-dependent Kinase Inhibitors in Human Gastric Carcinoma Cell Lines

Deregulation of cyclin, cyclin-dependent kinases (CDKs) and their inhibitors could have a pivotal role in the development of diverse human cancers. We examined the genetic status and the expression of CDK inhibitors ( p21, p27, pl6 and p15 ), CDK2 and cyclins (A, D1 and E) in eight gastric carcinoma cell lines, in comparison with the status of p53 gene alterations. All the cell lines (except MKN-28) that contained a p53 gene abnormality expressed very low or undetectable levels of p21 mRNA, while the cell lines (MKN-45 and -74) with wild-type p53 gene expressed high levels of p21 mRNA. An inverse correlation was found between the level of p21 mRNA and the expression of mRNAs for CDK2 and G1 cyclins. MKN-28 was an exception; it contained mutated p53 , and expressed mRNAs for p21 , CDK2 and G1 cyclins at high levels. Only MKN-45 and -74, with wild-type p53 , expressed considerable levels of p21 protein. Homozygous deletion of the p16 and p15 genes was detected in two (MKN-45 and HSC-39) of the eight gastric carcinoma cell lines. p16 protein was not expressed in three cell lines (MKN-28, MKN-74 and KATO-III), as well as MKN-45 and HSC-39. Rearrangement of the p15 gene was found in TMK-1. Rearrangement of the p27 gene was detected in MKN-45, although the expression of p27 protein was well preserved in all the gastric carcinoma cell lines. The expression of pRb was also preserved in all the cell lines except KATO-III. No obvious correlation was observed between the p53 gene status and the expression of p27 and p16 . These findings suggest that abnormal regulation of CDK2/cyclins and CDK inhibitors might be involved in deregulated growth of gastric carcinomas.     

DNA methyltransferase inhibitor 5-aza-CdR enhances the radiosensitivity of gastric cancer cells

The National Comprehensive Cancer Network guidelines recommend radiotherapy as a standard treatment for patients with a high risk of recurrence in gastric cancer. Because radiation is harmful to the surrounding organs, a radiation sensitizer might therefore be useful to decrease the side effects of patients with advanced gastric carcinoma. The aim of the current study was to clarify the effect of a DNA methyltransferase inhibitor, 5-aza-2'-deoxycytidine (CdR), on radiation sensitivity in gastric cancer cells. Five gastric cancer cell lines, OCUM-2M, OCUM-12, KATO-III, MKN-45, and MKN-74, were used. The effects of 5-aza-CdR with irradiation on the growth activity, cell-cycle distribution, apoptosis, and apoptosis-associated gene expression were examined. 5-aza-CdR sensitized three of five gastric cancer cell lines to radiation. A combination of irradiation and 5-aza-CdR significantly ( P <  0.05) decreased the growth activity compared with irradiation alone in OCUM-2M, OCUM-12, and MKN-45 cells, but not in KATO-III and MKN-74 cells. The percentage of cells in G₂–M phase and the apoptotic rate with irradiation in combination with 5-aza-CdR were increased in OCUM-2M, OCUM-12, and MKN-45 cells compared with irradiation alone, but not in KATO-III and MKN-74 cells. 5-aza-CdR increased the expression of p53 , RASSF1 , and death-associated protein kinases (DAPK) genes compared with the control or irradiation alone. These findings suggest that 5-aza-CdR might therefore be useful as a radiation sensitizer to treat some types of gastric carcinoma. The arrest at G₂–M phase and increased apoptotic rate might be partly mediated by enhanced expression of the p53 , RASSF1 , or DAPK gene families by 5-aza-CdR. ( Cancer Sci 2009; 100: 181–188)     

Krüppel-like factor 4 negatively regulates β-catenin expression and inhibits the proliferation, invasion and metastasis of gastric cancer

Krüppel-like factor 4 (KLF4) is a zinc-finger protein that plays an important role in the progression of gastric carcinoma. The abnormal activation of β-catenin frequently occurs in gastric cancer and has been associated with the promotion of tumor growth, invasion and metastasis. However, the potential interaction between KLF4 and β-catenin during gastric cancer development is unknown. In this study, a lentiviral KLF4 expression vector was constructed and utilized to transfect the human gastric cancer cell lines, SGC-7901, BGC-823, MKN-28 and MKN-45. KLF4 and β-catenin expression levels were measured by quantitative real-time RT-PCR and western blot analysis. Cell proliferation, colony formation and invasive potential were determined in the KLF4-transfected gastric cancer cells. The expression of E-cadherin and matrix metallopeptidase 2 (MMP2) was determined by western blot analysis. The overexpression of KLF4 significantly inhibited the expression of β-catenin in the MKN-45 gastric cancer cells. The restored expression of KLF4 suppressed proliferation, colony formation and inhibited the invasion and metastatic properties of MKN-45 gastric cancer cells. Furthermore, the forced expression of KLF4 in gastric tumor cell lines restored E-cadherin expression and inhibited MMP2 expression. Consistent with the in vitro findings, the enforced expression of the KLF4 gene in MKN-45 gastric carcinoma cells by lentiviral vector-mediated gene transfer effectively suppressed tumor growth in vivo. Our results show that KLF4 inhibits β-catenin expression and regulates the β-catenin-mediated biological behaviors of gastric cancer cells. The modulation of KLF4 expression may represent a novel therapeutic approach for β-catenin-driven malignancies.     

Sema3A在胃癌中的表达及意义

目的:探讨Sema3A在胃癌中的表达及意义。方法:利用酶联免疫吸附法（ELISA）检测正常健康人及胃癌患者血清中Sema3A表达的差异性。应用实时荧光定量聚合酶链反应(Real time PCR)检测胃癌细胞系、人正常胃黏膜细胞中Sema3A mRNA的表达差异。应用免疫印迹法（Western Blot)检测胃癌细胞系、人正常胃黏膜细胞中Sema3A 蛋白的表达差异。结果:与正常健康对照者相比,Sema3A在胃癌患者血清中表达下降（P<0.001）,且与胃癌患者的年龄、肿瘤分化程度相关（P<0.05）。人转移潜能胃癌细胞株MKN-28M、MKN-28NM中Sema3A mRNA和蛋白表达量明显低于人正常胃黏膜细胞（P<0.05）,且在高转移潜能MKN-28M细胞中表达量最低（P<0.05）。结论:与正常对照组相比,Sema3A在胃癌患者血清、人胃癌细胞株中均表达下降,且在胃癌高转移潜能MKN-28M细胞中表达最低,提示Sema3A在胃癌的发生发展中具有重要作用,可能扮演着抑癌基因的作用。     

乳腺癌细胞中AIP1/YAP/TAZ的表达及临床意义

目的:通过检测乳腺癌4T1细胞系中AIP1/YAP/TAZ的表达量及敲除AIP1后YAP/TAZ表达量的差异,为乳腺癌寻找新的生物标志物和治疗手段。方法:选取乳腺癌4T1细胞系,通过RT-PCR及Western-Blot检测AIP1/YAP/TAZ的表达量及其差异。结果:AIP1/YAP/TAZ在乳腺癌中均有表达,且敲除AIP1后YAP表达量明显减少（P<0.05）,TAZ表达量也减少。结论:AIP1/YAP/TAZ可作为一种潜在的生物标志物来预测乳腺癌的恶性程度,为探索乳腺癌的治疗提供新的方向。     

4-1BB基因在不同分化胃癌细胞株中的表达

目的：研究共刺激因子4-1BB在人胃高分化腺癌细胞株MKN-28、人胃中分化腺癌细胞株SGC-7901、人胃低分化腺癌细胞株BGC-823、人胃粘液腺癌细胞株MGC-803中的表达及其生物学意义。方法：用RT-PCR法测定胃癌细胞株中4-1BB mRNA表达;免疫细胞化学法测定胃癌细胞株中4-1BB蛋白表达;MTT法测定人淋巴细胞对胃癌细胞的体外杀伤活性;ELISA法检测人淋巴细胞与胃癌细胞共培养上清液中IL-2、IL-10、TNF-α的含量。结果：RT-PCR结果示4-1BB mRNA表达率从高到低依次为：MGC-803（77.30%）、BGC-823（71.68%）、SGC-7901（40.06%）、MKN-28（37.65%）。免疫细胞化学结果显示4-1BB着色程度由深到浅依次为：MGC-803、BGC-823、SGC-7901、MKN-28。MTT结果显示各时间段（24、48、72h）不同效靶比（10∶1,20∶1,40,∶1）下,人淋巴细胞对胃癌细胞的体外杀伤活性由高到低的顺序为：MKN-28、SGC-7901、BGC-823、MGC-803。ELISA结果显示在各效靶细胞比下,人淋巴细胞与胃癌细胞共培养上清液中IL-2、TNF-α的含量由高到低的顺序均为：MKN-28、SGC-7901、BGC-823、MGC-803。而IL-10的含量由高到低的顺序为：MGC-803、BGC-823、SGC-7901、MKN-28。结论：人胃癌细胞株MKN-28、SGC-7901、BGC-823、MGC-803中4-1BB基因的mRNA和蛋白均有表达,且胃癌细胞株的恶性程度越高其4-1BB表达水平越高;淋巴细胞对低表达4-1BB胃癌细胞的杀伤力较高;提示4-1BB可能通过抑制细胞因子TNF-α、IL-2和促进细胞因子IL-10的产生调节肿瘤免疫。     

In-vitro effect of a combination of 5-fluorouracil (5-FU) and cisplatin (CDDP) on human gastric cancer cell lines: timing of cisplatin treatment

Abstract. Background: To date, we have few effective chemotherapeutic agents against advanced and recurrent gastric cancer. 5-Fluorouracil (5-FU) and cisplatin (CDDP) are the most widely used drugs, and their combination has demonstrated favorable outcomes. However, the method (especially the timing) of CDDP administration is not well established. Methods: We examined the in-vitro effect of a combination of 5-FU and CDDP on four human gastric cancer cell lines: MKN-1, MKN-28, MKN-45, and MKN-74. The cell lines were exposed to 50% of the inhibitory concentrations of 5-FU and CDDP for 72 h and 8 h, respectively. CDDP was applied before, simultaneously with, and after the start of treatment with 5-FU. Results: When CDDP was applied after 5-FU, the cytotoxic activity against MKN-28, MKN-45, and MKN-74 was significantly potentiated. Against MKN-1, the earlier the initiation of CDDP treatment, the stronger was the cytotoxic effect. Conclusion: Our results suggest that the cytotoxicity of a combination of 5-FU and CDDP against human gastric cancer cells is both cell-line- and schedule-dependent and is especially affected by the timing of the CDDP treatment. Received: August 22, 2001 / Accepted: December 7, 2001     

Expression of Interleukin-6 and Its Effect on the Cell Growth of Gastric Carcinoma Cell Lines

The expression and the effect of IL-6 were examined in human gastric carcinoma cell lines to determine whether IL-6 serves as a growth stimulator. The expression of IL-6 mRNA was detected in three (TMK-1, MKN-1, MKN-7) of 8 gastric carcinoma cell lines. All three cell lines secreted IL-6 into the culture fluid, in large amounts in the cases of MKN-1 and MKN-7 cells. Scatchard plot analysis of IL-6 binding revealed that MKN-1 and MKN-7 cells had both high- and low-affinity receptors. Cell growth of MKN-1 and MKN-7 cells was stimulated by IL-6, while anti-IL-6 antibody inhibited growth. The expression of IL-la mRNA by these three cell lines was induced by IL-6. IL-la increased the expression of mRNA for IL-6 by TMK-1 cells. These findings indicate that IL-6 induced by IL-la is an autocrine growth factor for some gastric carcinomas     

PAR1 participates in the ability of multidrug resistance and tumorigenesis by controlling Hippo-YAP pathway

The Hippo pathway significantly correlates with organ size control and tumorigenesis. The activity of YAP/TAZ, a transducer of the Hippo pathway, is required to sustain self-renewal and tumor-initiation capacities in cancer stem cells (CSCs). But, upstream signals that control the mammalian Hippo pathway have not been well understood. Here, we reveal a connection between the Protease-activated receptor 1 (PAR1) signaling pathway and the Hippo-YAP pathway in gastric cancer stem-like cells. The selective PAR1 agonist TFLLR-NH₂ induces an increase in the fraction of side population cells which is enriched in CSCs, and promotes tumorigenesis, multi cancer drug resistance, cell morphological change, and cell invasion which are characteristics of CSCs. In addition, PAR1 activation inhibits the Hippo-YAP pathway kinase Lats via Rho GTPase. Lats kinase inhibition in turn results in increased nuclear localization of dephosphorylated YAP. Furthermore, PAR1 activation confers CSCs related traits via the Hippo-YAP pathway, and the Hippo-YAP pathway correlates with epithelial mesenchymal transition which is induced by PAR1 activation. Our research suggests that the PAR1 signaling deeply participates in the ability of multi drug resistance and tumorigenesis through interactions with the Hippo-YAP pathway signaling in gastric cancer stem-like cells. We presume that inhibited YAP is a new therapeutic target in the treatment human gastric cancer invasion and metastasis by dysregulated PAR1 or its agonists. The Hippo pathway significantly correlates with organ size control and tumorigenesis. The activity of YAP/TAZ, a transducer of the Hippo pathway, is required to sustain self-renewal and tumor-initiation capacities in cancer stem cells (CSCs). But, upstream signals that control the mammalian Hippo pathway have not been well understood. Here, we reveal a connection between the Protease-activated receptor 1 (PAR1) signaling pathway and the Hippo-YAP pathway in gastric cancer stem-like cells. The selective PAR1 agonist TFLLR-NH₂ induces an increase in the fraction of side population cells which is enriched in CSCs, and promotes tumorigenesis, multi cancer drug resistance, cell morphological change, and cell invasion which are characteristics of CSCs. In addition, PAR1 activation inhibits the Hippo-YAP pathway kinase Lats via Rho GTPase. Lats kinase inhibition in turn results in increased nuclear localization of Dephosphorylated YAP. Furthermore, PAR1 activation confers CSCs related traits via the Hippo-YAP pathway, and the Hippo-YAP pathway correlates with epithelial mesenchymal transition which is induced by PAR1 activation. Our research suggests that the PAR1 signaling deeply participates in the ability of multi drug resistance and tumorigenesis through interactions with the Hippo-YAP pathway signaling in gastric cancer stem-like cells. We presume that inhibited YAP is a new therapeutic target in the treatment human gastric cancer invasion and metastasis by dysregulated PAR1 or its agonists.     

Histone deacetylase inhibitor, trichostatin A, increases the chemosensitivity of anticancer drugs in gastric cancer cell lines

Epigenetic alterations of the histone acetylation play an important role in the regulation of gene expression associated with cell cycles and apoptosis that may affect the chemosensitivity of gastric carcinomas. Recently, a histone deacetylase inhibitor, trichostatin A (TSA), was proven to be a chemo-sensitizer on human erythroleukemia cells. With the aim of improving the chemotherapeutic efficacy of gastric carcinoma, the effect of TSA on the chemosensitivity of several anticancer drugs in gastric carcinoma cells was investigated. Human gastric cancer cell lines, OCUM-8 and MKN-74, and 5 anticancer drugs, 5-fluorouracil (5-FU), paclitaxel (PTX), oxaliplatin (OXA), irinotecan (SN38) and gemcitabine (GEM) were used. In both gastric cancer cell lines, a synergistic anti-proliferative effect by the combination of TSA (30 ng/ml) with 5-FU, PTX or SN38 showed a synergistic anti-proliferative effect in OCUM-8 and MKN-74 cells. TSA increases the expression of p21, p53, DAPK-1 and the DAPK-2 gene in both OCUM-8 and MKN-74 cells. In conclusion, TSA is a promising chemotherapeutical agent in combination with anticancer drugs of 5-FU, PTX and SN38 in gastric cancer cell lines. The up-regulation of p53, p21, DAPK-1 and DAPK-2 might be associated with the synergistic effect of TSA.     

Expression of cyclooxygenase-2 in dysplasia of the stomach and in intestinal-type gastric adenocarcinoma.

PURPOSE: Cyclooxygenase (Cox) is the key enzyme in conversion of arachidonic acid to prostanoids. Two Cox genes have been cloned, and expression of Cox-2 mRNA and protein has been shown to be elevated in several human malignancies and in animal models of carcinogenesis. The purpose of this study was to investigate Cox-2 protein expression in human gastric dysplasias and adenocarcinomas. EXPERIMENTAL DESIGN: Performance of several Cox-2 antibodies was evaluated, after which Cox-2 protein expression was studied in 67 gastric cancer specimens and in eight definitive dysplasias by using immunohistochemistry. RESULTS: Cox-2 positivity was detected in 58% (25/43) of the intestinal-type (well-differentiated) tumors and 6% (1/18) of diffuse-type (poorly differentiated) tumors. Consistent with these data, we detected higher expression of Cox-2 mRNA, protein, and enzymatic activity in well-differentiated gastric cancer cell lines (MKN-28 and MKN-74) when compared with poorly differentiated cell lines (HSC-39 and KATO III). Cox-2 immunoreactivity was localized to the carcinoma cells, but the stroma of the tumors was negative. However, strong Cox-2 positivity was consistently detected in stromal cells at sites of erosions and ulcerations. Furthermore, four of nine (44%) definitive dysplasias of the stomach that showed no evidence of invasion were positive for Cox-2. CONCLUSIONS: Cox-2 is expressed by the neoplastic cells in the intestinal-type gastric adenocarcinoma and by precarcinogenic (dysplastic) lesions leading to this disease.     

AJUBA调控MST1 YAP1和TAZ因子促进食管鳞状细胞癌细胞体外增殖 迁移和侵袭

  目的  探讨AJUBA在食管鳞状细胞癌（esophageal squamous cell carcinoma,ESCC）细胞中表达以及是否通过调控MST1、YAP1和TAZ因子影响细胞增殖、侵袭和迁移。  方法  采用Western blot 法检测AJUBA 蛋白在食管癌细胞系KYSE30、KYSE150与KYSE450中的表达水平,构建shRNA 干扰载体AJUBA转染至KYSE150食管癌细胞系;通过体外克隆形成实验、流式细胞术、划痕实验和Transwell实验探究AJUBA对KYSE150细胞的增殖、周期、迁移和侵袭能力的影响。采用RT-PCR和Western blot检测MST1、YAP1和TAZ mRNA和蛋白的表达;核质分离实验检测AJUBA、YAP1和TAZ蛋白表达情况。  结果  稳定干扰AJUBA基因后,平板克隆实验结果显示,与阴性对照组相比,shAJUBA组的细胞克隆形成数量明显减少,差异具有统计学意义（P<0.001）;流式细胞周期实验结果显示,shAJUBA组中的细胞阻滞于G₀/G₁期,G₂/M期与S期细胞显著减少（P<0.05）;划痕实验和Transwell侵袭实验结果显示,与AJUBA空染组相比,shAJUBA组细胞迁移能力和侵袭能力均明显减弱（P<0.001）;RT-PCR与Western blot实验结果显示,shAJUBA组细胞中的MST1、YAP1、TAZmRNA与蛋白表达水平均显著降低,差异具有统计学意义（P<0.05）。核质分离实验结果提示AJUBA蛋白在正常食管鳞状上皮细胞株（SHEE）细胞核中表达,而在KYSE150细胞胞质与胞核中表达;YAP1和TAZ蛋白在SHEE细胞中不表达,在KYSE150细胞质与细胞核中表达,上述结果差异均具有统计学意义（均P<0.05）。  结论  AJUBA促进食管癌细胞的增殖、侵袭与迁移,可能与激活MST1、YAP1、TAZ因子的表达而影响食管鳞状细胞癌进展有关。