Slug Inhibits Proliferation of Cervical Cancer Cells Through Down-regulating CDH3/β-catenin/C-myc Expression / 肿瘤防治研究

Objective To investigate the related mechanism of Slug inhibiting the proliferation of cervical cancer cell through CDH3/β-catenin/C-myc. Methods SiHa cells with stable Slug expression were screened. The expression of CDH3 in Slug-overexpressed SiHa cell was detected by RNA-sequence, Real-time PCR, Western blot and immunocytochemistry. The expression of CDH3 in SiHa and HeLa cells were detected by Western blot and immunocytochemistry. The protein level of CDH3 was up-regulated in HeLa cells or rescued in SiHa-Slug cells by transient transfection of CDH3 expression vector. The protein levels of β-catenin and C-myc were detected by Western blot, the cell growth was detected by cell counting and CCK-8 assays. Luciferase reporter assay and chromatin immunoprecipitation assay (ChIP) were performed to detect the effect of Slug on regulating the promoter region of CDH3. Results SiHa cell line with stable Slug expression was successfully constructed. Slug overexpression inhibited CDH3 expression in SiHa cells. CDH3 promoted cell proliferation and up-regulated the protein level of β-catenin and C-myc in HeLa and SiHa-Slug cells. Slug could recognize and bind to the E-boxes in the CDH3 promoter region and inhibited the transcription of CDH3 in SiHa cells. Conclusion Slug could inhibit the expression of β-catenin and C-myc by inhibiting CDH3 transcription in SiHa cells, and then attenuate the growth of SiHa cells.

Notch1 mediates breast cancer epithelial-mesenchymal transition through Slug / 西安交通大学学报(医学版)

Objective: To investigate the mechanism of Nocth1 in regulating epithelial-mesenchymal transition (EMT) in breast cancer. Methods: Jagged1 or shRNA was applied to regulate Notch signaling expression. EMT process and migration and invasion in breast cancer were determined. Moreover, Slug luciferase reporter assay was used to investigate the effect of Notch1 on Slug expression. And Slug expression was regulated by shRNA. The effects of Slug on Notch1 mediated breast cancer EMT process and migration and invasion were determined. Results: Jagged1 activated Notch signaling and promoted EMT process and migration and invasion in breast cancer. Overexpression of Notch1 could induce Slug promoter activation, and Slug overexpression could reverse the inhibition of Notch1 shRNA on breast cancer's EMT process and migration and invasion. Conclusion: Notch1 regulates EMT process, migration and invasion in breast cancer through targeting the Slug promoter.

Expressions of epithelial-mesenchymal transition-associated genes in human hepatocellular carcinoma cells / 西安交通大学学报(医学版)

Objective: To investigate the expressions of molecular markers associated with epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma (HCC) cells so as to lay foundation for determining related HCC cells model. Methods: We detected the mRNA and protein expression levels of EMT-associated genes, including E-cadherin, ZO-1, N-cadherin, Vimentin, Snail and Slug in five HCC cell lines, using quantitative real-time polymerase chain reaction analysis, Western blot and immunofluorescence. Results: The mRNA of EMT-associated genes was expressed in all of the five cell lines. The expressions of E-cadherin, ZO-1, N-cadherin and Slug protein were detected in all of the five cell lines. Vimentin protein was found in SNU368, SNU739, HLF and MHCC97L cells. Snail protein was detected in SNU368, SNU739, Huh-1 and MHCC97L cells. Conclusion: HLF seems to display the phenotype of mesenchymal cells whereas Huh-1 and SNU368 remain the epithelial phenotype.

Expression of E-cadherin, Slug and NCAM and its relationship to tumor invasiveness in patients with acromegaly

Pituitary adenomas account for 10-15% of primary intracranial tumors. Growth hormone (GH)-secreting adenomas account for 13% of all pituitary adenomas and cause acromegaly. These tumors can be aggressive, invade surrounding structures and are highly recurrent. The objective of this study was to evaluate E-cadherin, Slug and neural cell adhesion molecule (NCAM) expression in GH-secreting pituitary adenomas and its relationship to tumor invasiveness. A cross-sectional study of patients who underwent hypophysectomy due to GH-secreting pituitary adenoma from April 2007 to December 2014 was carried out. The medical records were reviewed to collect clinical data. Immediately after surgery, tumor samples were frozen in liquid nitrogen and stored in a biofreezer at -80°C for assessment of E-cadherin 1 (CDH1), SLUG (SNAI2), and NCAM (NCAM1) by real-time PCR. The samples were fixed in formalin and embedded in paraffin for immunohistochemical analysis of E-cadherin and NCAM. Thirty-five patients with acromegaly were included in the study. Of these, 65.7% had invasive tumors. Immunohistochemically, E-cadherin was expressed in 96.7% of patients, and NCAM in 80% of patients. There was no statistically significant relationship between tumor grade or invasiveness and immunohistochemical expression of these markers. Regarding gene expression, 50% of cases expressed CDH1, none expressed SNAI2, and 53.3% expressed NCAM1. There was no statistically significant relationship between tumor grade or invasiveness and gene expression of CDH1, SNAI2, and NCAM1. The absence of Slug overexpression and of E-cadherin and NCAM suppression suggests that expression of these markers is not associated with tumor invasiveness in GH-secreting pituitary adenomas.

Expressions of Slug, BRAF V600E and STIP1 proteins and their correlation with capsular invasion and regional lymph node metastasis in papillary thyroid carcinoma / 肿瘤研究与临床

Objective To observe the expressions of Slug, BRAF V600E and STIP1 proteins in papillary thyroid carcinoma (PTC), and to explore their correlation with capsular invasion and regional lymph node metastasis. Methods Slug, BRAF V600E and STIP1 expressions in 107 cases of differentiated PTC were examined by immunohistochemical staining. The expressions of three proteins and clinicopathological data were statistically analyzed. Results Positive rates of Slug, BRAF V600E and STIP1 in PTC were 65.4 % (70/107), 61.7 % (66/107) and 66.4 % (71/107), respectively, and overexpression of Slug, BRAF V600E and STIP1 was significantly associated with capsular invasion and regional lymph node metastasis in PTC (P< 0.05). There are a significant correlation between expression of Slug and BRAF V600E in PTC (r= 0.235, P< 0.05). Conclusion Overexpression of Slug, BRAF V600E and STIP1 proteins is associated with capsular invasion and regional lymph node metastasis in PTC, which maybe useful for predicting regional lymph node metastasis and prognostic evaluation.

Detection of KIT gene of a case with serious phenotypes of piebaldism / 中山大学学报(医学科学版)

[Objective]To investigate the mutations of KIT gene and SLUG(SNAI2)gene in one patients with piebaldism in Chi?na.[Methods]All coding exons and exon-intron boundaries of KIT gene and SLUG gene were amplified by PCR. The PCR products were sequenced. The DNA samples from 50 normal subjects were also sequenced for control.[Results]The novel mutation,c.860T>A (p.V287E),was detected in patient. This mutation was absent in his parents and the controls ,indicating a de novo mutation. The de?tection result of all coding exons and exon-intron boundaries of SLUG gene was normal.This p.V287E mutation was located in the ex?tracellular ligand-bindingdomain(ectodomain)of KIT,which may generate clash with E249 and disrupt the conformation ofβD andβD/βE of D3 that required for SCF(stem cell factor)binding.[Conclusion]We have identified a novel mutation of KIT gene,c.860T>A(p.V287E),which is probably associated with serious phenotypes of piebaldism.

Expressions of epithelial-mesenchymal transition-associated genes in cervical cancer cell lines / 西安交通大学学报(医学版)

Objective Epithelial-mesenchymal transition (EMT)is thought to be one of the important mechanisms of invasion and metastasis of carcinoma cells,which includes down-regulation of epithelial cell markers and up-regulation of mesenchymal markers.The aim of our study was to investigate whether EMT occurs in cervical cancer cell lines and determine the proper cell model to study the mechanism of EMT in cervical cancer.Methods We detected the mRNA expression of EMT-associated genes,namely,E-cadherin,vimentin,Snail and Slug,in four cervical cancer cell lines using reverse transcriptase-polymerase chain reaction analysis.E-cadherin and vimentin protein expression levels were examined using Western blotting. Snail and Slug protein expression levels were examined by immunocytochemical staining.Results Vimentin and Snail mRNA was expressed in all of the four cell lines.E-cadherin and Slug mRNA was expressed in SiHa,HeLa and RJC-1 cells except CS1213 cells.E-cadherin protein was found in SiHa and RJC-1 cells whereas vimentin protein was detected in HeLa and CS1213.Snail protein was expressed in all of the four cell lines.Slug protein was found in SiHa,HeLa and RJC-1 cells.Conclusion It seems that HeLa and CS1213 display the phenotype of mesenchymal cells whereas SiHa and RJC-1 remain the epithelial phenotype.

Effects of CTHRC1 on human ovarian cancer cell metastasis and its mechanism / 上海交通大学学报（医学版）

Objective·To investigate the role of collagen triple helix repeat containing-1 (CTHRC1) in ovarian cancer cell metastasis and the related mechanism. Methods·The expression of CTHRC1 in ovarian cancer cells was detected by Western blotting. The cell line which had high expression of CTHRC1 was transfected with a CTHRC1 specific shRNA, and the lenti-CTHRC1 was used to overexpress CTHRC1 in the cell line whose expression of CTHRC1 was very low. Then the expression of Slug and MMP-2 was assessed. Transwell assay was used to determine the migration and invasion capability of ovarian cancer cells after transfection. Results·The expression of CTHRC1 in HO8910 cells was the lowest, while the CTHRC1 protein level was dramatically increased after transfection of lenti-CTHRC1. Meanwhile, there was a distinct rise of the migration and invasion ability, as well as the expression of Slug and MMP-2 (all P<0.05). Conversely, CaOV3 cells had a higher protein expression of CTHRC1. By using lenti-shCTHRC1, a remarkable knockdown of CTHRC1 was obtained. Likewise, the capability of migration and invasion was decreased, and the Slug and MMP-2 expression was reduced (all P<0.05). Conclusion·CTHRC1 might positively regulate Slug and MMP-2 to promote ovarian cancer cell metastasis.

Expression and significance of Slug in papillary thyroid carcinomas / 实用医学杂志

Objective To explore the clinical significance of Slug expression in papillary thyroid carci-noma(PTC). Methods Employed Ventana immunohistochemistry assay to determine the expression of Slug in 107 cases of PTC and para-tumorous normal tissue. The relationship with Slug expression in PTC and clinico-pathology data were also analyzed. Results Expression of Slug in PTC (65.4%, 70/107)and para-tumorous nor-mal tissue (14.0%,15/107)were statistically different (P < 0.001). Overexpression of Slug in PTC was signifi-cantly associated with capsular invasion and lymph node metastasis (P < 0.05). Conclusion Overexpression of Slug in PTC is associated with capsular invasion and lymph node metastasis , these may suggest some clinical significance of Slug expression in PTC in predicting lymph node metastasis and prognosis.

Effects of Emodin on Migration and Invasion Ability and Expressions of E-cadherin and Slugof Human Hepatoma Cell Lines HepG2 / 中国中医药信息杂志

Objective To investigate the effects of emodin on the migration and invasion ability and expressions of E-cadherin and Slug of human hepatoma cell lines HepG2; To discuss the possible mechanisms of anti-hepatocellular carcinoma.Methods HepG2 cells were cultured in vitro. The experimental group was treated with emodin at concentrations of 10μmol/L and 20μmol/L as. The negative control group was treated with the same volume of RPMI-1640 medium, while the positive control group was treated with 10μmol/L floxuridine. The cell matrix adhesion assay, wound healing and transwell chamber in vitro invasion assay were used to observe the effects of emodin on HepG2 cell adhesion rate and migration and invasion ability. Western blot analysis was used to observe the changes of expressions of E-cadherin and Slug.Results Compared with the negative control group, emodin inhibited significantly HepG2 cell adhesion rate and migration and invasion ability were in a dose-dependent manner (P<0.05,P<0.01); Western blot analysis showed that the protein expression of E-cadherin increased significantly, and the level of Slug decreased significantly in a dose-dependent manner (P<0.05,P<0.01).Conclusion Emodin can significantly inhibit migration and invasion of HepG2 cells, which mechanism may up-regulate expressions of E-cadherin and down-regulate Slug.

Tolfenamic Acid Inhibits the Proliferation, Migration, and Invasion of Nasopharyngeal Carcinoma: Involvement of p38-Mediated Down-Regulation of Slug

PURPOSE: Tolfenamic acid (TA), a non-steroidal anti-inflammatory drug, is known to exhibit antitumor effects in various cancers apart from nasopharyngeal cancer (NPC). NPC exhibits high invasiveness, as well as metastatic potential, and patients continue to suffer from residual, recurrent, or metastatic disease even after chemoradiation therapy. Therefore, new treatment strategies are needed for NPC. In this study, we investigated the efficacy and molecular mechanisms of TA in NPC treatment. MATERIALS AND METHODS: TA-induced cell death was detected by cell viability assay in the NPC cell lines, HNE1 and HONE1. Wound healing assay, invasion assay, and Western blot analysis were used to evaluate the antitumor effects of TA in NPC cell lines. RESULTS: Treatment with TA suppressed the migration and invasion of HNE1 and HONE1 cells. Hepatocyte growth factor enhanced the proliferation, migration, and invasion abilities of NPC cells. This enhancement was successfully inhibited by TA treatment. Treatment with TA increased phosphorylation of p38, and the inhibition of p38 with SB203580 reversed the cytotoxic, anti-invasive, and anti-migratory effects of TA treatment in NPC cell lines. Moreover, inhibition of p38 also reversed the decrease in expression of Slug that was induced by TA treatment. CONCLUSION: In conclusion, the activation of p38 plays a role in mediating TA-induced cytotoxicity and inhibition of invasion and migration via down-regulation of Slug.

The expression and significance of Slug,EMMPRIN and E-cadherin in salivary adenoid cystic carcinoma / 实用口腔医学杂志

Objective:To investigate the expression of Slug,EMMPRIN and E-cadherin in salivary adenoid cystic carcinoma (SACC)and its correlation with clinicopathological characteristics,and the correlation among themselves.Methods:Slug,EMMPRIN and E-cadherin expression in 1 1 5 SACC cases of SACC was examined by immunohistochemical staining.The results and clinicopatho-logical data were statistically analyzed.Results:High positive expression frequencies of Slug(76.5%)and EMMPRIN(69.6%)and low positive expression frequency of E-cadherin(51 .3%)were found in 1 1 5 SACC cases.The expression of Slug and EMMPRIN was positively associated with the histopathological types,clinical stages,perineural invasion,recurrence and distance metastasis(P <0.05).The expression of E-cadherin was negatively associated with the histopathological types,clinical stages,perineural invasion and distance metastasis(P <0.05).There was a significant correlation between Slug and EMMPRIN expression(P <0.05),negative correlation between EMMPRIN and E-cadherin expression(P <0.05)and between Slug and E-cadherin expression(P <0.05).Con-clusion:The expression of Slug,EMMPRIN and E-cadherin is closely correlated to the clinicopathological characteristics of SACC.

Down regulation of miR-203 in radiation-induced thymic lymphoma promoted cells proliferation and inhibited apoptosis / 中华放射医学与防护杂志

Objective To investigate the role of miR-203 in radiation-induced thymic lymphoma (RITL).Methods A 60Co irradiator was used for total-body irradiation.MicroRNAs(miRNAs) level was assayed by qRT-PCR.Cell proliferation was assayed by MTT assay.Cell apoptosis was examined by fluorescence activated cell sorter (FACS).Dual luciferase reporter assay system was used to detect the 3'UTR reporter.Results MiR-203 was down-regulated in RITL tissues.Overexpression of miR-203 strongly inhibited the proliferation of both NIH3T3 cells and EL4 cells and vice versa.MiR-203 inhibited cells proliferation and induced apoptosis via TANK-binding kinase (TBK1),SLUG (SNAI2) and Cyclin D1 (CCND1).Conclusions Radiation down-regulated the level of miR-203 in thymic,which promoted radiation-induced thymic lymphoma by targeting TBK1,SNAI2 and CCND1.

miR-200c regulates migration of breast cancer cell BT549 by targeting Slug / 中国免疫学杂志

Objective:To investigate the effect on the expression of Slug for the trasfection of miR-200c combined with the research on the ability of migration of breast cancer cell BT549.Methods:Chemically synthesized miR-200c mimic was trasfected into BT549 cells,which have high metastatic potential.The effect on the ability of migration of breast cancer cell BT549 for the transfection of miR-200c was analyzed by Transwell migration assay and Wound healing assay.The expression of Slug and E-cadherin mRNA was detected by Real-time PCR.The expression of Slug protein was detected by Western blot.Results:Transfection with miR-200c mimic significantly down-regulated the expression of Slug as compared with the control group (P<0.05).BT549 cell tranfected with miR-200c mimic had lower levels of migration capacity than cells in the control group (P<0.05).Conclusion:miR-200c inhibits Epithelial-mes-enchymal transition by suppressing Slug leading to down-regulation of migration capacity of breast cancer cell BT549.

First record of qualitative losses caused by Meghimatium pictum in vineyards of Southern Brazil and the effects of two molluscicides for its control / Primeiro Registro de perdas qualitativas causadas por Meghimatium pictum em vinhedos do Sul do Brasil e os efeitos de dois moluscicidas para o seu controle

This research reports the terrestrial slug Meghimatium pictum (Stoliczka, 1873) (Stylommathophora: Philomycidae) as an agricultural pest for the first time in Southern Brazil vineyards. The species was found in densities exceeding 20 slugs.m-2 in Vitis labrusca L. vineyards at six municipalities of the Southern Brazil's viticulture region. It causes damage a loss by leaving residual mucus on grapes and by consuming grapes already perforated by other organisms, such as insects or birds, or mechanically damaged by in situ compression. The effectiveness of iron phosphate and metaldehyde baits on M. pictum was evaluated in laboratory experiments. Iron phosphate bait was more effective in controlling M. pictum (70%) than metaldehyde bait (15%).

O presente trabalho relata pela primeira vez a lesma terrestre Meghimatium pictum (Stoliczka, 1873) (Stylommathophora: Philomycidae) como uma praga agrícola causando danos em vinhedos no Sul do Brasil. Esta espécie foi encontrada em densidades superiores a 20 lesmas.m-2 danificando uvas da espécie Vitis labrusca L. em seis municípios da região vitícola do Sul do Brasil. O impacto econômico causado por M. pictum está associado ao movimento dos espécimes no dossel da videira e nas uvas, provocando a contaminação residual por muco e o consumo de uvas já perfuradas por outros organismos, tais como insetos ou aves, ou mecanicamente danificadas pela compressão in situ. A eficácia das iscas a base de fosfato de ferro e metaldeído sobre M. pictum também foi avaliada em condições de laboratório. A isca a base de fosfato de ferro foi mais eficiente no controle de M. pictum (70%) do que a isca a base de metaldeído (15%).

Knockdown of Erk/Slug signals increases radiosensitivity of MDA-MB-231 cells to γ-rays by upregulating PUMA / 中华放射医学与防护杂志

Objective To study the role Erk/Slug signal pathway in the radiosensitivity of MDAMB-231 cells.Methods MDA-MB-231 cells were transfected with NF-κBp65 siRNA,PUMA siRNA and Slug siRNA respectively or treated with U0126 for 24h,then the cells were irradiated with 4 Gy γ-ray.At different time points post-irradiation,the expressions of Erk1/2,NF-κBp65,Slug and PUMA protein were detected.The cell survival rate and apoptotic index were detected by MTT and TUNEL methods.Resluts Compared with 4 Gy irradiated group,the expression of PUMA was reduced in NF-κB p65 siRNA/4 Gy group,the expressions of Slug and Erk1/2 were obviously decreased but PUMA increased in U0126/4 Gy group,the expression of Erk1/2 had no change but the expression of PUMA increased significantly in Slug siRNA/4 Gyγ group.Meanwhile,at 48h post-irradiation,for U0126/4 Gy group and Slug siRNA/4 Gy group,cells survival rates were decreased to 19.78 ±2.71 (F=11.39,P＜0.05) and 17.41 ±4.58(F=15.31,P＜0.05),cell apoptosis rates were 28.61 ±4.70 (F=9.84,P＜0.05) and 27.55±6.41(F =10.31,P ＜ 0.05),respectively.At 24 h post-irradiation,for NF-κB p65 siRNA/4 Gy group and PUMA siRNA/4 Gy group,cell survival rates approached to 85.65 ± 9.60 (F =12.31,P ＜ 0.05) and 87.53±11.50 (F=13.68,P＜0.05),and cell apoptosis rates declined to 3.28 ±0.78 (F=10.83,P ＜ 0.05) and 3.46 ± 0.84 (F =9.92,P ＜ 0.05).Conclusions The radiosensitivity of MDA-MB-231cells was relative to the induction of NF-κB up-regulated PUMA,and the radioresistance was caused by the up-regulation of Slug induced by Erk1/2,which inhibited the expression of PUMA.

Effect of Slug knock-down on tumor formation and metastasis of HCT116 cells in nude mice / 中国免疫学杂志

Objective:To explore the influence of RNAi mediated Slug silencing on the growth and metastasis of colon cancer in nude mice.Methods: HCT116 colon cancer cells use for 24 five-week-old nude mice implanted subcutaneously , established colon cancer xenograft model in nude mice ,all divided into blank control group ,negative control group and the experimental group ,each group had eight nude mices.All group were injected with saline , negative plasmid and lentivirus vectors respectively.Tumor growth was observed and draw tumor curved growth ,changes in tumor growth and lymph node metastasis between the groups were observed ,Slug gene and protein expression were detected by immunohistochemistry ,qRT-PCR and Western blot analysis.Results: Slug gene shRNA intervention group compared with the control group and negative control group ,tumor grew slower ,tumor mass was significantly reduced (3.08±0.31 vs 7.37±1.18,7.46±1.16,P＜0.01),experimental group of lymph node-positive rate was 36.3%( 4 /11 ) ,compared to the negative control group 77.8% ( 14/18 ) and the control group was 68.4% ( 13/19 ) ( P＜0.01 ).Conclusion: Targeted Slug RNA interference can significantly inhibit the growth of colon cancer in nude mice ,lymph node metastasis and the expression of the gene protein in cancer tissue ,Slug may be a potential molecular target for colon cancer gene therapy.

Comparative Analysis for Estimating Hydraulic Conductivity Values to Improve the Estimation of Groundwater Recharge in Yaoundé-Cameroon.

Climate change will deeply affect the precipitation and evapotranspiration around the world. The sustainability of groundwater resources is crucial for regional and local communities, which is intimately tied to the changing recharge rate. To accurately assess the recharge rate, different methods were used to estimate hydraulic conductivity of an unconfined aquifer in this study. Particle size method with four empirical formulae, together with in-situ aquifer tests and the inverse modelling techniques were integrated to evaluate their potential for the determination of hydraulic conductivity of unconsolidated aquifer materials in order to improve groundwater recharge estimation. Results showed a wide disparity between the granulometric estimates of the hydraulic conductivity and the in-situ and modelling techniques. Slug test values range from 5.13 x 10-6 m/s to 4.96 x 10-5 m/s whereas the infiltration test (Porchet method) results vary from 1.91 x 10-7 m/s to 1.16 x 10-6 m/s. The simulated hydraulic conductivity values range from 2.54 x 10-7 m/s to 6.36 x 10-7 m/s, with a decreasing trend in the northeast-southwest (NE-SW) direction. The infiltration method appeared to be better than the granulometric one in the estimation of the vertical hydraulic conductivity within the unsaturated zone of porous formations. This study also pointed out that within an anisotropic formation, the hydraulic conductivity ratio (Kv/Kh) should not always be taken as equal to 10. Specific tests should be implemented to access this value in a given aquifer.The inverse modelling results showed the net recharge values varying from 68.5 mm/yr to 180 mm/yr. The modelling technique appears to be consistent with the in-situ estimates. Therefore, the application of groundwater modelling tool in this study has shown excellent promise for characterizing the spatial distribution of hydraulic conductivity and net recharge values within the targeted aquifer system.

Expression of Slug in pancreatic cancer and inhibitory effects of anti-Slug on invasion and migration of pancreatic carcinoma cells / 中华肝胆外科杂志

Objective To investigate expression of slug and E-cadherin in pancreatic cancer tissues and determine the inhibitory effects of anti-Slug, an anti-sense plasmid, on the invasion of pancreatic cancer cell lines in vitro. Methods Slug and E-cadherin protein and mRNA was analyzed by IHP and RT-PCR in 36 cases of pancreatic cancer. Then anti-Slug plasmid was transfected into herin and Slug expression. The inhibitory effects of anti-sense Slug were also detected by Transwell motility assay and Matrigel invasion assay. Results The expression of Slug and mRNA in metastatic pancreatic cancer tissue was higher than that in non-metastatic tissue. E-cadherin and mRNA was lower in metastasis tissues(P＜0.05). The inverse relationships were further observed by transient transfection of anti-Slug into SW1990H4 cells. The downregulated expression of Slug and re-expression of E-cadherin were found. The Slug mRNA levels were 0.985±0.016,0.973±0.014, 0. 554±0. 011 after 0, 48 h of transfection of anti-sense Slug, and that of E-cadherin were 0.120±0.001, 0.360±0.002, 0. 727±0. 006, respectively. The diference was significant between different time points (P＜0.05). The Slug mRNA levels were 0. 206±0.017, 0.968±0.015, and that of E-cadherin were 0. 18±0.002,0.727±0.006 after stable transfection of anti-sense Slug, and control plasmid, respectively. The diference was significant (P＜0.05). The motility activity(393±28, 352±24, 96 ±13 )and the invasion activity (223 ± 69, 202 ± 64, 65 ±19) of1 antisense Slug transfectant cells were significantly decreased as compared with those of control cells (P＜0.05). Conclusions Higher expression of slug and lower expression of E-cadherin is related to the invasion and metastasis in pancreatic cancer. A reverse corelation of E-cadherin and Slug expression exists in pancreatic cancer. Slug is possibly a potential target for cancer gene therapy blocking invasion and metastasis in human pancreatic cancer.

Radiosensitivity of AsPC-1 cell to γ-rays enhanced by up-regulation of PUMA induced by targeted Slug gene / 中华放射医学与防护杂志

Objective To explore the influence of PUMA on radiosensitivity of pancreatic cancer AsPC-1 cells after Slug gene inhibition by transfected short interferencing RNA(siRNA). Methods The AsPC-1 cells were infected with MOI 10,50,100 for 72 h, respectively. The expression of Slug and PUMA was analyzed by Western blotting and immunohistochemistry methods. The transfected and control cells were exposed to 4 Gy γ-rays. The cells inhibition rate was examined by MTT, Hoechst 33342 and IP double staining. DNA ladder and Giemsa staning was used to observe apoptosis. Results The relative value of Slug expression was 0.831 ±0.14,0. 546 ±0.12 and 0.178 ±0.08 after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly lower than that of control group ( F = 4. 992,P ＜ 0.05 ).The relative value of PUMA was 0. 325 ±0. 07,0. 593 ±0. 11 and 0. 978 ±0. 12, after AsPC-1 was infected with Slug-siRNA ( MOI 10,50,100) for 72 h, significantly higher than that of control group ( F = 4. 324,P ＜ 0. 05 ). The cell proliferation rate was ( 78.76 ± 9. 36 ) % in transfection combined with radiosensitivity group, significantly higher than that of transfection group [ ( 43.68 ± 6.71 ) % ] and radiosensitivity group alone [( 19.25 ± 3.72)% ] (F = 5.056, P ＜ 0.05). The apoptosis of transfection combined with radiosensitivity group was significantly higher than that of others. Conclusions Slug gene targeting siRNA could inhibit the expression of Slug, and consequently increase the activation of PUMA expression, and so enhance the radiosensitivity to γ-rays.