BACKGROUND/AIMS: Transforming growth factor-β1 (TGF-β1) induction of epithelial-mesenchymal transition (EMT) is one of the mechanisms by which colorectal cancer (CRC) cells acquire migratory and invasive capacities, and subsequently metastasize. Parthenolide (PT) expresses multiple anti-cancer and anti-inflammatory activities that inhibit nuclear factor κB by targeting the IκB kinase complex. In the present study, we aimed to investigate whether PT can inhibit TGF-β1-induced EMT in CRC cell lines.METHODS: HT-29 and SW480 cell lines were used in the experiment. Cell viability was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and sub-G1 analysis was measured by flow cytometry. The induction of EMT by TGF-β1 and inhibition of the process by PT was analyzed by phase contrast microscopy, wounding healing, cellular migration and invasion assays, and Western blotting.RESULTS: TGF-β1 inhibits HT-29 cell proliferation, but has no effect on SW480 cell proliferation; different concentrations of TGF-β1 did not induce apoptosis in HT-29 and SW480 cells. PT attenuates TGF-β1-induced elongated, fibroblast-like shape changing in cells. PT inhibits TGF-β1-induced cell migration and cell invasion. In addition, other EMT markers such as β-catenin, Vimentin, Snail, and Slug were suppressed by PT, while E-cadherin was increased by PT.CONCLUSIONS: Our findings show that PT inhibits TGF-β1-induced EMT by suppressing the expression of the mesenchymal protein and increasing expression of the epithelial protein. These findings suggest a novel approach for CRC treatment by suppression of TGF-β1-induced EMT.
Apoptosis ; Blotting, Western ; Cadherins ; Cell Line ; Cell Movement ; Cell Proliferation ; Cell Survival ; Colorectal Neoplasms ; Epithelial-Mesenchymal Transition ; Flow Cytometry ; Gastropoda ; HT29 Cells ; Humans ; Microscopy, Phase-Contrast ; Phosphotransferases ; Snails ; Transforming Growth Factors ; Vimentin ; Wounds and Injuries

p90 ribosomal S6 kinase (p90RSK), one of the downstream effectors in ERK1/2 pathways, shows high expression in human breast cancer tissues. However, its role in breast cancer development and drug resistance is not fully understood. Here, we demonstrate that Cis-DDP treatment failed to increase cytotoxicity in MDA-MB-231 cells compared to MCF-7 cells and p90RSK activation was involved in Cis-DDP-resistance to MDA-MB-231 cells. In the study, we found that inhibition of p90RSK expression or activation using a small interfering RNA (siRNA) or dominant-negative kinase mutant (DN-p90RSK) plasmid overexpression increased Cis-DDP-induced cytotoxicity of MDA-MB-231 cells, respectively. Mechanistically, we found that Cis-DDP resistance was associated with up-regulation of epithelial growth factor (EGF) expression and EGF treatment induced cancer survival signaling pathway including activation of ERK1/2, p90RSK, and Akt. We also examined the expression of epithelial-mesenchymal transition (EMT)-associated proteins using a reverse transition-quantitative PCR analysis. Cis-DDP treatment induced EMT by increasing the expression levels of N-cadherin, Snail, and Twist, while decreasing the expression levels of E-cadherin. Furthermore, we examined the epithelial marker, Zonula occludens-1 (ZO-1) using immunofluorescence analysis and found that Cis-DDP-inhibited ZO-1 expression was recovered by p90RSK deactivated condition. Therefore, we conclude that Cis-DDP resistance is involved in EMT via regulating the EGF-mediated p90RSK signaling pathway in MDA-MB-231 cells.
Breast Neoplasms ; Cadherins ; Cisplatin ; Drug Resistance ; Epidermal Growth Factor ; Epithelial-Mesenchymal Transition ; Fluorescent Antibody Technique ; Humans ; MCF-7 Cells ; Phosphotransferases ; Plasmids ; Polymerase Chain Reaction ; Ribosomal Protein S6 Kinases, 90-kDa ; RNA, Small Interfering ; Snails ; Triple Negative Breast Neoplasms ; Up-Regulation

The small GTP-binding protein Rab25 is associated with tumor formation and progression. However, recent studies have shown discordant effects of Rab25 on cancer cell progression depending on cell lineage. In the present study, we elucidate the underlying mechanisms by which Rab25 induces cellular invasion. We demonstrate that Rab25 increases β1 integrin levels and subsequent activation of EGFR and upregulation of VEGF-A expression, leading to increased Snail expression, epithelial-to-mesenchymal transition and cancer cell invasiveness. Strikingly, we identify that Snail mediates Rab25-induced cancer cell invasiveness through fascin expression and that ectopic expression of Rab25 aggravates metastasis of ovarian cancer cells to the lung. We thus demonstrate a novel role of a β1 integrin/EGFR/VEGF-A/Snail signaling cascade in Rab25-induced cancer cell aggressiveness through induction of fascin expression, thus providing novel biomarkers and potential therapeutic targets for Rab25-expressing cancer cells.
Biomarkers ; Cell Lineage ; Ectopic Gene Expression ; GTP-Binding Proteins ; Lung ; Neoplasm Metastasis ; Ovarian Neoplasms ; Snails ; Up-Regulation ; Vascular Endothelial Growth Factor A

PURPOSE: Cancer-associated fibroblasts (CAFs) activated by cancer cells has a central role in development and malignant biological behavior in colorectal cancer (CRC). Adult fibroblasts do not express Snail, but Snail-positive fibroblasts are discovered in the stroma of malignant CRC and reported to be the key role to chemoresistance. However, the reciprocal effect of CAFs expressed Snail to chemoresistance on CRC cells and the underlying molecular mechanisms are not fully characterized. MATERIALS AND METHODS: Snail-overexpressed 3T3 stable cell lines were generated by lipidosome and CT26 mixed with 3T3-Snail subcutaneous transplanted CRC models were established by subcutaneous injection. Cell Counting Kit-8, flow cytometry and western blotting assays were performed, and immunohistochemistry staining was studied. The cytokines participated in chemoresistance was validated with reverse transcriptase-polymerase chain reaction and heatmap. RESULTS: Snail-expression fibroblasts are discovered in human and mouse spontaneous CRCs. Overexpression of Snail induces 3T3 fibroblasts transdifferentiation to CAFs. CT26 co-cultured with 3T3-Snail resisted the impairment from 5-fluorouracil and paclitaxel in vitro. The subcutaneous transplanted tumor models included 3T3-Snail cells develop without restrictions even after treating with 5-fluorouracil or paclitaxel. Moreover, these chemoresistant processes may be mediated by CCL1 secreted by Snail-expression fibroblasts via transforming growth factor β/nuclear factor-κB signaling pathways. CONCLUSION: Taken together, Snail-expressing 3T3 fibroblasts display CAFs properties that support 5-fluorouracil and paclitaxel chemoresistance in CRC via participation of CCL1 and suggest that inhibition of the Snail-expression fibroblasts in tumor may be a useful strategy to limit chemoresistance.
Adult ; Animals ; Blotting, Western ; Cell Count ; Cell Line ; Colorectal Neoplasms* ; Cytokines ; Drug Resistance, Multiple ; Fibroblasts* ; Flow Cytometry ; Fluorouracil ; Humans ; Immunohistochemistry ; In Vitro Techniques ; Injections, Subcutaneous ; Mice ; Paclitaxel ; Snails ; Transforming Growth Factors

PURPOSE: The biological function of long non-coding RNAs (lncRNAs) is only partially understood; therefore, in this study, we investigated the expression of the novel HOXA11 antisense (HOXA11as) lncRNA and its oncogenic role in serous ovarian cancer (SOC). MATERIALS AND METHODS: HOXA11as expression was examined in 129 SOC tissue samples by real time reverse transcription polymerase chain reaction. Clinicopathological factors and patient survival were compared between the high (n=27) and low HOXA11as expression group (n=102). To investigate the role of HOXA11as in cell proliferation, invasion, and migration, HOXA11as expression in ovarian cancer cells was knocked down using RNA interference. RESULTS: HOXA11as expression in cancer tissue was 77-fold higher than that of noncancerous tissue (p < 0.05). Higher HOXA11as expression was significantly correlated with histological grade (p=0.017) and preoperative cancer antigen 125 (p=0.048). HOXA11as overexpression in SOC cells led to increased cell proliferation, invasion, and migration. Moreover, HOXA11as was associated with the expression of genes involved in cell invasion, migration, and epithelial-mesenchymal transition (EMT), including vascular endothelial growth factor, matrix metalloproteinase 9 (MMP-9), B-catenin, E-cadherin, Snail, Twist, and vimentin. Multivariate analysis revealed that HOXA11as was a prognostic factor of progressive disease and mortality (hazard ratio [HR], 1.730; p=0.043 and HR, 2.170; p=0.033, respectively). Progression-free and overall survival were significantly shorter in patients with high HOXA11as expression. CONCLUSION: These findings highlight the clinical significance of HOXA11as to predicting the prognosis of SOC patients and suggest its potential in promoting tumor aggressiveness via regulation of vascular endothelial growth factor (VEGF), MMP-9, and EMT-related mechanisms.
Cadherins ; Cell Proliferation* ; Epithelial-Mesenchymal Transition ; Humans ; Matrix Metalloproteinase 9 ; Mortality ; Multivariate Analysis ; Ovarian Neoplasms* ; Polymerase Chain Reaction ; Prognosis* ; Reverse Transcription ; RNA Interference ; RNA, Long Noncoding* ; Snails ; Vascular Endothelial Growth Factor A ; Vimentin

Fasciola hepatica is a trematode that causes zoonosis mainly in cattle and sheep and occasionally in humans. Fascioliasis has been reported in Korea; however, determining F. hepatica infection in snails has not been done recently. Thus, using PCR, we evaluated the prevalence of F. hepatica infection in snails at 4 large water-dropwort fields. Among 349 examined snails, F. hepatica-specific internal transcribed space 1 (ITS-1) and/or ITS-2 markers were detected in 12 snails and confirmed using sequence analysis. Morphologically, 213 of 349 collected snails were dextral shelled, which is the same aperture as the lymnaeid snail, the vectorial host for F. hepatica. Among the 12 F. hepatica-infected snails, 6 were known first intermediate hosts in Korea (Lymnaea viridis and L. ollula) and the remaining 6 (Lymnaea sp.) were potentially a new first intermediate host in Korea. It has been shown that the overall prevalence of the snails contaminated with F. hepatica in water-dropwort fields was 3.4%; however, the prevalence varied among the fields. This is the first study to estimate the prevalence of F. hepatica infection using the vectorial capacity of the snails in Korea.
Animals ; Base Sequence ; DNA, Helminth/chemistry/genetics ; DNA, Ribosomal Spacer/chemistry/genetics ; Fasciola hepatica/anatomy & histology/genetics/*isolation & purification ; Molecular Sequence Data ; Oenanthe/growth & development ; *Polymerase Chain Reaction ; Republic of Korea ; Sequence Analysis, DNA ; Snails/growth & development/*parasitology

OBJECTIVETo investigate the epidemiological situation of cercarial trematodes infection in freshwater snails from different water resources in Chiang Mai province, Thailand.

METHODSThe snail specimens were collected from 13 districts of Chiang Mai province during April 2008 to February 2012. The prevalence of cercarial infection in snails was investigated using the crushing method. The drawing was done with the help of a camera lucida for the morphological study.

RESULTSA total of 2 479 snail individuals were collected and classified into 7 families, 11 genera, and 14 species, Among them, 8 snails species were found to be infected with an overall prevalence of 17.27% (428/2 479), which infected with nine groups of cercariae; gymnocephalous cercaria, strigea cercaria, megalurous cercaria, monostome cercaria, parapleurolophocercous cercaria (Haplorchis cercaria), pleurolophocercous cercaria, furcocercous cercaria (Transversotrema cercaria), xiphidiocercaria, and virgulate cercaria. The parapleurolophocercous cercaria was found to be the dominant type among the cercarial infection in the snails (64.25%).

CONCLUSIONSThe various species of snails found in the research location act as the intermediate hosts for the high prevalence of parasitic infection of many species of mammals. This work will provide new information on both the distribution and first intermediate host of trematodes.

Animals ; Cercaria ; growth & development ; isolation & purification ; Fresh Water ; Prevalence ; Snails ; parasitology ; Thailand ; epidemiology ; Trematoda ; growth & development ; isolation & purification

We detected metacercariae of Echinostoma revolutum in Filopaludina sp. snails purchased from a local market in Nam Dinh Province for the first time in Vietnam. Adult flukes were harvested from experimentally infected hamsters at days 14 and 17 post-infection. The metacercariae were round, 170-190 microm (n=15) in diameter, with a cyst wall thickness of about 12 microm. A total of 37 collar spines were arranged around the head collar, and large excretory granules were seen in 2 canals of the excretory bladder. The 14-day old adult flukes were elongated, ventrally curved, and 5.0-7.2x0.8-1.3 mm (n=20). The head collar had a total of 37 collar spines arranged in 2 alternating rows, including 5 corner spines on each side. The cirrus sac contained a saccular seminal vesicle, a prostatic gland, and an unarmed cirrus. Two tandem testes were smooth or slightly lobed. Eggs were ovoid to elliptical, 110-118x70-75 microm. These morphological characters were similar to those of E. revolutum and E. jurini. We tentatively identified it as E. revolutum because the validity of E. jurini remains to be elucidated. The taxonomic relationship of E. revolutum and E. jurini is discussed.
Animals ; Cricetinae ; Echinostoma/anatomy & histology/classification/growth & development/*isolation & purification ; Echinostomiasis/*parasitology ; Female ; Metacercariae/anatomy & histology/classification/growth & development/isolation & purification ; Snails/*parasitology ; Vietnam

OBJECTIVETo research molluscicidal effect activity, active components and stable passage of endophyte JJ18 from Pseudolarix amabilis and examine the possibility for practical application.

METHODMolluscicidal effect test was performed according to the immersion test method suggested by WHO.

RESULTImmersion test with JJ18 broth showed that the active components were extracellular moiety of the broth and that 10% concentration solution could kill nearly 90% snail immersed after 72 h, the salified broth has favourable thermostabily and photostability and showed that JJ18 has stable passage and its active components concentrate in the extract of n-butanol.

CONCLUSIONThe metabolite of endophyte JJ18 has activity for molluscicidal effect and potential for application.

Animals ; Bacteria ; chemistry ; growth & development ; Bacterial Proteins ; pharmacology ; Molluscacides ; chemistry ; pharmacology ; Pinaceae ; microbiology ; Polysaccharides, Bacterial ; pharmacology ; Snails ; drug effects

OBJECTIVETo construct the life cycle of Angiostrongylus cantonensis (A.cantonensis) in laboratory condition.

METHODSSD rats were infected orally with the third-stage larvae of A.cantonensis collected from Jiangmen, Guangdong province. Six weeks after infection, the first-stage larvae were isolated from fresh feces of the rats by using Baermann funnel to infect 25 second-generation white jade snails raised in laboratory at the daily dose of 300 000 for 3 consecutive days. Three weeks later, the snails were dissected for counting the third-staged larvae of A.cantonensis, and those positive for A.cantonensis infection were fed directly to 10 fasting rats. The serum samples of the rats were then collected 2 weeks later for examination of specific antibodies using ELISA. The feces of the infected rats were examined microscopically after 6 weeks, and the brain, heart and lungs of the infected rats were dissected to observe the larvae at 3, 5, and 8 weeks, respectively.

RESULTSThe 3-stage larvae of A.cantonensis were found in the second-generation snails 3 weeks after infection. The positivity rate of serum specific antibodies was 100% in the 10 rats 2 weeks after feeding of the infected snails. The 1-stage larvae were detected in the feces of the rats 6 weeks after infection, and the fourth-stage larvae were found in the brain of the rats at 3 weeks, while adult worm and eggs were found in the heart and lungs of the infected rats at 5 and 8 weeks.

CONCLUSIONThe successful establishment of human colon carcinoma cell line with PRL-3 gene knock-down provide a basis for investigation of the role of PRL-3 gene in the metastasis of human colorectal carcinoma.

Angiostrongylus cantonensis ; growth & development ; physiology ; Animals ; Disease Vectors ; Larva ; growth & development ; physiology ; Life Cycle Stages ; Rats ; Rats, Sprague-Dawley ; Rodent Diseases ; parasitology ; Snails ; parasitology