Snail Research Articles

The effects and mechanism of IL-6 on the epithelial to mesenchymal transition of pancreatic cancer cells

Objective To investigate the effects and mechanism of IL-6 on the epithelial to mesenchymal transition of human pancreatic cancer cells.Methods IL-6 was added into the culture media of human pancreatic cancer cells Capan-2,SW1990,and STAT3-siRNA-SW1990.Cell growth was measured by MTT assays.STAT3,p-STAT3,Snail,Twist,and E-cadherin mRNA and protein expression were examined using real-time fluorescence quantitative polymerase chain reaction (RT-PCR)and Western blot,respectively.The invasion abilities of SW1990 and Capan-2 cells were determined by a cell invasion assay in vitro.Results Our results showed that 100 μg/L of IL-6 significantly promoted the growth and invasion abilities of Capan-2 and SW1990 cells (P＜0.05).The use of IL-6 not only markedly increased the protein expression of P-STAT3 and Snail,but also greatly decreased the mRNA and protein expression of E-cadherin.The use of IL-6 can not change the mRNA and protein expression of Snail and E-cadherin.Conclusion Activation of the STAT3 signal transducer pathway with IL-6 can promote the epithelial to mesenchymal transition of pancreatic cancer cells in vitro through up-regulation of Snail and down-regulation of E-cadherin expression.Therefore the STAT3 signal transducer may provide a novel therapeutic target for the treatment of pancreatic cancer. Key words: Pancreatic neoplasms; Interleukin-6; Signal transduction; Epithelium

MORPHOLOGICAL AND ETHOLOGICAL ASPECTS OF MATING IN TWO SPECIES OF THE FAMILY HELICIDAE (GASTROPODA PULMONATA): THEBA PISANA (MÜLLER) AND HELIX APERTA BORN

SUMMARY The mating behaviour of two Helicidae (Gastropoda Pulmonata) species, Theba pisana (Muller) and Helix (Cantareus) aperta Born, is described. Comparison with prior studies suggests that the position assumed for mating is not determined by the ecology of the species but is an occasionally acquired element. Each partner reacts to the stimuli of the other. The stimuli are necessary for the initiation of the approach and the realization of a correct mating position and appear to induce one snail to accept the other. If one of the subjects is not sufficiently motivated the stimuli are ineffective. The stimuli are not specific and responses indicating a simple cause-effect relationship were not observed. The shooting of the dart marks the beginning of attempts at copulation and occurs when, after repeated stimulation, a threshold of excitement is exceeded. In T. pisana and H. aperta, although the darts are shot simultaneously after the contact organs have been pressed closely together, this happens only ...

CD133 promotes the invasion and metastasis of gastric cancer via epithelial-mesenchymal transition

To examine the association between CD133 expression and invasion of gastric cancer, and to elucidate whether CD133 can promote the invasion and metastasis of gastric cancer via epithelial-mesenchymal transition (EMT). The CD133(+) and CD133(-) KATO-III( cells were sorted by magnetic activated cell sorting (MACS). The invasion ability was detected by Transwell method. RT-PCR and Western blot were used to detect the expression of EMT-related factors in KATO-III( cells before and after CD133 was knocked out by siRNA method. The expressions of CD133 and EMT-related proteins of cancer and adjacent normal tissues in 50 patients with gastric cancer were detected by Western blot, and correlations among protein expressions were also analyzed. As compared to CD133(-) cells, the number of broken-membrane cells was significantly higher (67.7±10.5 vs. 13.3±6.8, P=0.001) and the invasion ability was stronger (P<0.05) in CD133(+) cells, while the mRNA expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells (0.311±0.015 vs. 0.223±0.016, P=0.040; 0.581±0.020 vs. 0.270±0.018,P=0.004), and the protein expression levels of Snail and N-cadherin were significantly higher in CD133(+) cells as well (0.513±0.015 vs. 0.179±0.023, P=0.030; 0.538±0.028 vs. 0.202±0.032, P=0.020), but E-cadherin mRNA and protein levels were significantly lower in CD133(+) cells (0.231±0.009 vs. 0.460±0.015, P=0.040; 0.426±0.030 vs. 0.748±0.027, P=0.040). After CD133 knock-out, the expressions of Snail and N-cadherin were down-regulated (P<0.05) and the expression of E-cadherin was up-regulated (P<0.05). As compared to normal mucosal tissues, the protein expression levels of Snail, N-cadherin and CD133 in gastric cancer tissues were significantly higher(0.635±0.119 vs. 0.485±0.116, P=0.029; 0.599±0.114 vs. 0.259±0.108, P=0.020; 0.754±0.154 vs. 0.329±0.134, P=0.001), while the protein expression of E-cadherin in gastric cancer tissues was lower (0.378±0.123 vs. 0.752±0.156, P=0.003). The protein expressions of Snail and N-cadherin were positively correlated with CD133 expression (r=0.278, P=0.048; r=0.406, P=0.003) and the protein expression of E-cadherin was negatively correlated with CD133 expression (r=-0.504, P=0.000). CD133(+) cells in primary lesion of gastric cancer have relatively higher invasion ability, which may promote the metastasis of gastric cancer via up-regulation of EMT-related factors.

Expression of special AT rich sequence binding protein-1 in bladder transitional cell carcinoma and its correlation with epithelial-mesenchymal transition

Objective To investigate the expression of special AT rich sequence binding protein-1 (SATB1) and genes involved in epithelial-mesenchymal transition (EMT) in bladder transitional cell carcinoma (BTCC),and to explore the relationship between SATB1 and EMT-related genes with with clinieopathologic features of BTCC.Methods Immunohistochemisty and real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) were used to detect the expression of SATB1,E-cadherin and Snail in 82 cases of BTCC tissues.For each specimen,the cancerous tissue,peri-cancer tissue and its remote normal mueosa were analyzed and compared Results The immunohistochemisty revealed that the expression of SATB1 and Snail in BTCC was significantly higher than that in periearcinomatous tissues and normal bladder tissues (P ＜ 0.05).Inversely,the expression of E-cadherin was significantly lower in BTCC than that in pericarcinonatous tissues and normal bladder tissues (P ＜ 0.05).The positive expression rate of SATB1,E-cadherin and Snail was 90.24％,18.29％ and 80.49％ for BTCC,17.07％,79.27％ and 30.49％ for pericarcinomatouse tissues,and 8.54％,91.46％ and 15.85％ for normal bladder tissues,respectively.The positive expression rate of SATB1,E-cadherin and Snail in the pathological grades 1,2and3 was 76.19％,23.81％ and 61.90％,92.00％,20.00％ and 76.00％,and 97.22％,13.89％and 94.44％ respectively.There were significant differences in each group (P ＜ 0.05).The positive expression rate of SATB1 and Snail in the clinical stages of Tis-T1 was lower than that in the stages of T2-T4 (78.00％ vs.97.96％,P ＜ 0.05 ; 66.67％ vs.89.80％,P ＜ 0.05).Inversely,the positive expression rate of E-cadherin was significantly higher in the clinical stages of Tis-T1 than that in the stages of T2-T4 (24.24％ vs.14.29％,P ＜ 0.05).Moreover,with the advancing grades and appearances of lymph node metastasis,the expression levels of SATB1 and Snail were increased gradually,but in contrast,the expression levels of E-cadherin decreased gradually.There were significant differences in each group (P ＜0.05).The results of RT-qPCR data were agreed to those of the immunohistochemisty.Conclusion The excessive expression of SATB1 and Snail,and low expression of E-cadherin were closely related to the pathology,clinical stages and metastasis of lymph node of the cancer.SATB1 expression was inversely correlated to E-cadherin,but positively to Snail.SATB1 may regulate the invasion and metastasis of BTCC through inducing EMT.The over expression of SATB1 may play an important role in carcinogenesis and progress of BTCC. Key words: Bladder transitional cell carcinoma; Epithelial-mesenchymal transition; Special AT rich sequence binding protein-1 ; E-cadherin

In vivo phytotherapy of snail by plant derived active components in control of fascioliasis

Fascioliasis is worldwide zoonotic disease, which caused great economic loss to human being. In vivo killing of Fasciola larva (redia and cercaria) is one of the effective methods to control of fascioliasis. In vivo toxicity of different binary combinations (1:1ratio) of plant derived active components such as citral, ferulic acid, umbelliferone; azadirachtin and allicin were tested against Fasciola larva (redia and cercaria) in snail Lymnaea acuminata . Mortality of larvae were observed at 2h, 4h, 6h and 8h of treatment. In vivo exposure of azadirachtin + allicin (1:1 ratio) was highly toxic against redia and cercaria (8h LC 50 0.007and 0.006 mg/L). Combinations of citral + umbelliferone, cirtal + ferulic acid were less toxicity against redia and cercaria larva than azadirachtin + allicin. In vivo use of these larvicidal active components in the binary combinations at sublethal does will be more helpful in controlling the redia/cercaria than their individual components.

Attraction of eyestalk ablated Lymnaea acuminata towards the different photo and chemo stimulants

In the eye of Lymnaea acuminata cornea, lens, microvillar layer, pigmented layer, cell layer and photoreceptors works in well organized manner and respond to photic stimulation. In the present study attraction of eye stalk ablated snail Lymnaea acuminata towards the different visible monochromatic light was studied. The photosensitivity of snails to different monochromatic light was wavelength as well as time dependent. In left or right eye stalk ablated snails response to monochromatic light was noted after 3 and 7 days of ablation. However, when both eyes were ablated the response to photic stimulation was observed after 9 days. This functional recovery indicates that type and number of photoreceptors as well as their regenerative capacity are not same in left and right eye of uninfected and infected snails. Sixty minute exposure of red light+ serine as chemo stimulant in bait, caused maximum attraction of snails (62%) than single exposure of chemo (36%) or red (26%) light stimulus.

POPULATION DYNAMICS OF THE SNAIL THEBA PISANA (HELICIDAE) IN THE SAND DUNES OF NORTHERN ISRAEL

ABSTRACT The population dynamics of the land snail Theba pisana (Helicidae) was studied for two successive years in the sand dunes of northern Israel and under laboratory conditions. The field population in autumn consisted of two size classes: large snails of more than 10 mm in diameter and small snails of less than 5 mm in diameter. After the first rain, the large snails that emerged from aestivation laid eggs and died. After 3–4 weeks, a new cohort hatched and grew intensively throughout winter to the end of spring. However, a group of small snails ceased growing and entered an early aestivation. These growth-inhibited individuals retained immature size. Their proportion in the population is estimated at 5–10% throughout the year. The population in the laboratory showed a similar growth pattern and size distribution as that in the field. The inhibited-growth group in the laboratory was 40% of the population. We conclude that Theba pisana in Israel developed a strategy of mixed annual and biennial life ...

SELECTIVE LAND SNAIL PREDATION BY THE SPINY MOUSE, ACOMYS CAHIRINUS, IN NAHAL OREN, MT. CARMEL, ISRAEL

ABSTRACT Microclimatic differences between the south-facing slope and the north-facing slope of the Lower Nahal Oren, Mt. Carmel, Israel result in a dramatic biological divergence. The south-facing slope harbors predominantly African representatives, including the spiny mouse, Acomys cahirinus. The north—facing slope harbors predominantly European taxa, including the prosobranch land snail Pomatias olivieri. Here we show that predation by Acomys on European Pomatias exceeds the proportion of Pomatias shells among the large land snail fauna at the microsite. The ratios of consumed to unconsumed Pomatias were approximately 4:1 and 7:1 in the north—and south-facing slopes, respectively. In all other land snail species, the consumed constituted at most 2/3 of the unconsumed. Remarkably, the selective predation of Acomys on Pomatias is significantly higher on the warmer and drier south-facing slope. We hypothesize that natural selection led to an adaptive behavioral predator—prey relationship at the microsite.

STUDIES ON LARVAL STAGES OF DIGENETIC TREMATODES IN AQUATIC MOLLUSCS OF ISRAEL. V. THREE CERCARIAE FROM THE FRESHWATER SNAIL BITHYNIA SIDONIENSIS

ABSTRACT Three eercariae are described from the freshwater snail Bithynia sidoniensis Mousson, collected in the 'Ada stream (near Binyamina) in 1975; a furcocercaria of the Vivax type (Cercaria Levantina 14) and two xiphidiocercariae (Cercariae Levantinae 15 and 16). Details are presented on the morphology and swimming activities of the cercariae as well as on their development within and emergence from the snail. Encystation attempts by one of the cercariae (Cercaria Levantina 16) are described.

Effect of crowding on growth and mortality in the edible snail Helix lucorum (Gastropoda: Pulmonata) in Greece

ABSTRACT The effect of crowding on growth and mortality in Helix lucorum L. hatched in the laboratory was tested experimentally. The higher the number of grouped individuals, the smaller was the growth and the higher the mortality. The optimum density for this snail under the conditions studied seemed to be 50 individuals/m2 floor area. Densities observed in nature ranged 3–80 individuals/m2.

THE SAME GENE POOL IS SHARED BY TWO DISTINCT MORPHS OF THE FRESHWATER SNAIL MELANOPSIS FROM THE SEA OF GALILEE

ABSTRACT As a contribution to the taxonomy of the polymorphic snail Melanopsis in the Sea of Galilee, Israel, an electrophoretic survey based on 14 enzymes (15 putative loci) was carried out on about 900 snails, belonging to 2 distinct morphs (banded and unhanded), sampled at 17 locations from around the lake. Hedrick's (1971) measure of genotypic identity was used to calculate the genetic distances among the groups. According to a UPGMA phenogram, all groups belong to the same gene pool, with the shortest distances characterizing morphs collected at the same location.

SPERMATOGENESIS AND SPERM STRUCTURE OF THE NORMALLY PARTHENOGENETIC FRESHWATER SNAIL MELANOIDES TUBERCOLATA

ABSTRACT The freshwater snail Melanoides tuberculata ranges throughout Asia and Africa, and throughout much of its range it reproduces parthenogenetically. In Israel, however, males have been found. These males produce two types of spermatozoa, euspermatozoa and paraspermatozoa. The euspermatozoa are filiform (about 230 μ m in length) with a head comprising a nucleus and conical acrosome, a mid-piece (98 μ m long) of four equal-sized mitochondria, and a tail. Unlike in other cerithiaceans, the nucleus is unusually elongate (31 μ m long × 1 μ m diameter) and the axoneme is housed within an intranuclear canal. The paraspermatozoa consists of a head of electron-dense blocks (up to 25 μ m long) which surround up to 15 axonemes, a mid-piece of elongate mitochondria and a tail tuft, as in other cerithiaceans. Structural changes during spermatogenesis resemble those described for other gastropods with dimorphic spermatozoa.

Abiotic factors and anti-reproductive action of bait containing eugenol against Lymnaea acuminata.

The effect of abiotic factor (temperature, pH, dissolved oxygen, carbon dioxide) on the reproduction of snail L. acuminata fed to bait containing 40% and 80% of 24h LC50 eugenol have been reported in this paper. Feeding of baits containing sub-lethal concentration of eugenol caused a significant reduction in fecundity, hatchability and survival of young snails. Maximum (1644 egg/20 snails) and minimum (228 eggs/ 20 snails) fecundity was observed in month of May and December, respectively. There was a significant (P<0.05) change in the endogenous level of protein, amino acid, DNA and RNA in the ovotestis of L. acuminata. Treatment of 80 % of 24h LC50 with eugenol in bait caused maximum reduction in protein (52.12% of control), amino acid (11.89% of control), DNA (30.90% of control) and RNA (13.12% of control) level. Simultaneous, inhibition in acetylcholinesterase (AChE) activity in nervous tissue was also noted. Maximum inhibition in AChE activity (36.19% of control) was observed in snail exposed to 80 % of 24h LC50 of eugenol.

A new Brazilian tornid is possibly the flattest coiled snail

A new Brazilian tornid is possibly the flattest coiled snail

MiR-145 inhibits the lung adenocarcinoma initiating cells genesis via regulating epithelial-mesenchymal transition progress

Objective To investigate whether microRNA-145 (miR-145) could regulate the lung adenocarcinoma initiating cells genesis by using A549 cell line,and the possible molecular mechanisms.Methods The initiating cells microspheres were derived from the serum-free-cultured A549 celld.Pre-miR-145 mimics and anti-miR-145 inhibitor were transfected into A549 initiating cells microspheres by Lipofectamine 2000.The number of A549 initiating cell microspheres was counted by light microscopy,and the percent of CD133 + cells wsd measured by flow cytometry.Western blotting was applied for determining the protein levels of epithelial-mesenchymal transition (EMT) related proteins:Snail,Slug (EMT-related transcription factor),Cytokeratin 18,E-cadherin (epithelial markers) and N-cadherin,Vimentin (mesenchymal markers).Results As compared with the control group,the percent of CD133 + cells was remarkably decreased in Pre-miR-145 mimics groups [(25.39 ± 12.37) ％ vs (43.70 ± 17.58) ％,(P ＜0.05)],whereas significantly increased in the anti-miR-145 inhibitor groups [(58.89 ± 23.54) ％ vs (45.60 ± 18.58)％,(P ＜ 0.05)].The overexpression of miR-145 led to the downregulation of Snail,Slug,Vimentin and N-cadherin,but the upregulation of E-cadherin and Cytokeratin 18.However,repressing miR-145 apparently up-regulated the Snail,Slug,Vimentin and N-cadherin,and down-regulated the Ecadherin and Cytokeratin 18.Conclusion miR-145 can inhibit the A549 initiating cells genesis,which might be controbuted to the regulation of EMT pathway. Key words: Lung cancer; miR-145; Epithelial-mesenchymal transition

Spheroid cells derived from human colon cancer cells exhibit epithelial to mesenchymal transition characteristics

Objective To investigate the similarities between cancer stem cells and epithelial mes- enchymal transition （EMT） cells in spheroid cells. Methods Generate cancer spheroid cells from colon cancer cell line DLD-1 in serum free medium （SFM） and to investigate the expression of colon cancer stem cells markers such as CD133, CD166,1eucine-rich-repeat-containing G-protein-coupled receptor 5 （ Lgr5 ） and the numbers of CD133^＋ cells. In addition, the capacity of migration,invasion and tumorigenicity in vivo and in vitro were confirmed by transwell assay, colony formation assay and mouse xenograft model respec- tively. Moreover, the expression of EMT-related genes was detected by real-time quantitative polymerase chain reaction （Real-time PCR） and Western blotting. Results The spheroid cells exhibit higher migra- tion,invasion,colony-formation and tumorigenicity properties. The CD133^＋ subpopulation was increased in spheroid cells. Moreover, the colon cancer stem cell markers such as CD133, CD166, Lgr5 and mesenchymal genes such as Vimentin and Snail were increased while ZO-1 decreased in spheroid cells. Surprisingly, E- cadherin-the key mediator of EMT was higher expressed too. Conclusion Although the E-cadherin was in- creased expression, considered the experiments in vitro and in vivo, we thought that E-cadherin may be not the sole cause of the malignant files in spheroid cells. Moreover, the spheroid cells exhibit cancer stem cells nrooerties and EMT Characteristics. Key words: Colon cancer; Cancer stem cell; Epithelial mesenchymal transition

The impact of water level changes on the Oncomelania snail habitats in Poyang Lake regions before and after the impoundment of Three Gorges reservoir

To explore the impact of the impoundment of Three Gorges reservoir on Oncomelania snail habitats of Poyang Lake. Four counties with schistosomiasis endemic around the Poyang Lake, naming as Xingzi, Duchang, Jinxian and Xinjian, were selected as study areas. The average daily water level of four major hydrological stations near the above 4 counties from 1997 to 2008 were collected, as well as the elevation of 215 marshlands with snails in those 4 counties, the data of schistosomiasis endemic situation and snail survey of 4 counties in the year before the impoundment of Three Gorges reservoir (2002) and the 6th year after the impoundment of Three Gorges reservoir (2008). The fluctuations of water levels of Poyang Lake, flooding day of marshland with snails as well as the changes of the indicators in snail survey and prevention and control measures before and after the impoundment of Three Gorges reservoir were compared. Before the impoundment of Three Gorges reservoir, the fluctuations of average water level of the four hydrological stations around Poyang Lake was 6.91-12.93 m; which reduced to 5.72 - 10.75 m after the impoundment. The D-value of the fluctuations of average water level was (1.06 ± 0.47) m. The date of the annual maximum water level delayed by an average of 12 d; while the date of annual minimum water level arrived an average of 12 d earlier than the impoundment of Three Gorges reservoir. The average flooding days of marshland with snails were separately (100.04 ± 42.06) d before the impoundment and (64.13 ± 22.30) d after the impoundment. The difference was statistically significant (t = 3.70, P < 0.05). The density of snails in the 4 counties around Poyang Lake was 0.8437/0.1 m(2) after the impoundment; declining by 31.04% from the snail density (1.2234/0.1 m(2)) before the impoundment. The density of infectious snails was 0.0014/0.1 m(2) after the impoundment; dropping by 67.34% from it (0.0042/0.1 m(2)) before the impoundment; the infection rate of snails was separately 0.34% after the impoundment and 0.16% before the impoundment. The difference was statistically significant (χ(2) = 53.25, P < 0.05). The coverage rate of chemotherapy increased from 4.45% (35 558/798 200) in 2002 to 9.80% (79 457/810 400) in 2008; the difference was statistically significant (χ(2) = 17 336.63, P < 0.05). The coverage rate of snail killing increased from 1.35% (389.04/28 824.66) in 2002 to 7.49% (2260.90/30 166.22) in 2008; the difference was also statistically significant (χ(2) = 28 824.66, P < 0.05). The impact of the impoundment of Three Gorges reservoir shortened the flooding days of marshland with snails and helped the prevention and control of schistosomiasis in Poyang Lake regions as the environment went against the breeding of the snails.

Evaluation of Glucosamine and Snail Mucin on the Progression of Experimental Knee Osteoarthritis in Dogs

Se evaluaron los efectos de la glucosamina oral y la inyeccion intramuscular (IM) de mucina de caracol en la progresion de la osteoartritis (OA) experimental en perros. Fueron utilizados 20 perros mestizos adultos con un peso medio de 12,4±1,8 kg. La OA experimental se indujo quirurgicamente mediante el modelo de ranura. Los animales se dividieron aleatoriamente en tres grupos despues de la evidencia radiografica de OA. El grupo 1 (control, 10 perros) fue tratado con una solucion salina normal dos veces por semana durante cuatro semanas. El grupo 2 (5 perros) fue tratado con 10 mg/kg de glucosamina oral al dia por cuatro semanas, y el grupo 3 (5 perros) fue tratado con 5 mg/kg IM de una solucion de mucina de caracol al 5% dos veces por semana durante cuatro semanas. Se obtuvieron muestras de sangre desde la vena cefalica previo a la artrotomia quirurgica, despues de la artrotomia e inmediatamente despues de la confirmacion radiografica de OA (semana 0), y en el intervalo de dos semanas hasta cuatro semanas de tratamiento. La eficacia de los farmacos se evaluo por los cambios plasmaticos de IL-6 y MMP-3, mientras que la seguridad, se determino por los cambios en el volumen del hematocrito (VH), el recuento total de globulos blancos (RGB), y la observacion de reacciones adversas asociadas a la administracion de farmacos. El VH y RGB no difirieron significativamente (P>0,05) en el grupo control. Los niveles de IL-6 y MMP-3 plasmatica fueron significativamente mas bajas (P 0,05) en la IL-6 y MMP-3 entre los dos grupos de tratamiento. Ademas, se observo un edema doloroso en el sitio de inyeccion de los perros tratados con mucina de caracol. En los perros tratados con glucosamina oral no se observo reaccion adversa. Se concluye que tanto la glucosamina oral y la inyeccion IM de mucina de caracol modifican comparablemente la progresion de OA. Sin embargo, debido a la reaccion adversa observada con la inyeccion IM de mucina caracol, se necesitan estudios adicionales para determinar la via de administracion mas adecuada.

Enhanced antitumor effect of axitinib synergistic interaction with AG490 via VEGFR2/JAK2/STAT3 signaling mediated epithelial-mesenchymal transition in cervical cancer in vitro

Background: Epithelial to mesenchymal transition (EMT) not only confers tumor cells with a distinct advantage for metastatic dissemination, but also it provides tumor cells for proliferation and chemotherapy resistance. Thus, inhibiting this process using single or multiple agents, remains a field of intensive research. The development of EMT has been implicated in not only conferring cancer cells distant metastasis, but also providing tumor cells advantages for proliferation and chemotherapy resistance. Objective: We investigated whether axitinib synergistic interaction with AG490 could effectively block the growth and EMT-mediated tumor metastasis in cervical cancer in vitro. Methods: We addressed synergism effects on tumor growth, axitinib, AG490 alone and in any possible combinations on cell viability, apoptosis and cell cycle distribution were evaluated by XTT assay, Annexin V/propidium iodide staining and flow cytometric analysis. To assess how combination therapy affected chemotactic motility in tumorcells, wound-healing migration and the Boyden chamber invasion assays were applied in vitro. Further, using western blots, the key signaling molecules and pathways in response to axitinib and AG490 combination treatment in anti tumor growth and anti EMT-mediated tumor metastasis were analyzed in Hela cells. Results: Therapy with axitinib and AG490 resulted in strong synergistic inhibition of proliferation, migration and cell cycle arrest without potently effect on apoptosis induction relative to AG490 monotherapy and control in vitro. Furthermore, phosphorylation of VEGFR2 (Tyr1175), JAK2 (Tyr1007/1008) and STAT3 (Tyr705) was completelyblocked in parallel with significant reduction expressions of N-cadherin and obvious enhancement levels of E-cadherin through down-regulating Snail responding to combination treatment in cervical cancer cells. Conclusion:We first provided evidences that combination therapy of axitinib and AG490 significantly enhances anti-tumor effect via VEGFR2/JAK2/STAT3 signaling mediated growth arrest and EMT-mediated tumor metastasis inhibition, holds promise for an efficacious treatment of these advanced patients. Keywords: AG490, axitinib, EMT, Hela cells, JAK2, STAT3, VEGFR2

Ganesh Is Reading/The Snail

Ganesh Is Reading/The Snail