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Objective:To explore the clinical significance of the expression of nuclear factor of activated T cells 5 (NFAT5) in esophageal cancer tissues and the effect of the expression of knock-down esophageal cancer cells on their migration ability.Methods:The expression of NFAT5 in tissues of 26 patients with esophageal cancer and their adjacent tissues was detected by immunohistochemistry. Esophageal cancer cells ECA109 were divided into experimental group and control group. The experimental group ECA109 cells were transfected with NFAT5-siRNA plasmid, and the control group ECA109 cells were transfected with MOCK-siRNA plasmid. The mRNA content of NFAT5 was detected by fluorescence quantitative PCR. The expression of NFAT5, TLR4 and MyD88 proteins in the experimental group and control group were detected by Western blot. Transwell assay and cell scratch assay were used to detect the migration ability of the experimental group and the NC group.Results:Immunohistochemical test results showed that the positive rate of NFAT5 in esophageal cancer tissues was 80.77% (21 cases/26 cases) , and the expression rate was 15.38% (4 cases/26 cases) in corresponding adjacent tissues. The positive rate of NFAT5 protein in esophageal cancer tissues was significantly higher than that in adjacent tissues ( P<0.001) . The NFAT5 mRNA content of ECA109 cells in experimental group and control group decreased after transfection with corresponding siRNA. The protein expression levels of NFAT5, TLR4 and MyD88 in ECA109 cells in the experimental group were 0.28±0.08, 0.31±0.13 and 0.41±0.14, respectively. The protein expression levels of NFAT5, TLR4 and MyD88 in ECA109 cells in control group were 0.95±0.15, 0.84±0.22 and 1.04±0.26, respectively. The expression of TLR4 and MyD88 in esophageal cancer ECA109 cells decreased significantly ( P<0.05) . The scratch healing rate of esophageal cancer ECA109 cells was 52.67%±5.21% in the experimental group and 82.91%±7.26 % in the control group. Transwell experiment results showed that the number of successfully migrated cells in the experimental group was (35±5) , and the number of successfully migrated cells in the control group was (92±13) . The results showed that the migration ability of esophageal cancer ECA109 cells was significantly decreased after low expression of NFAT5 ( P<0.01) . Conclusions:The expression of NFAT5 is significantly increased in esophageal cancer tissues, and the expression of NFAT5 may be related to the malignant degree of esophageal cancer. Moreover, NFAT5 affects the migration ability of esophageal cancer cells by regulating TLR4/MyD88 signaling pathway.
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Objective:To investigate the expression of ras-related C3 botulinum toxin substrate 3 (RAC3) in glioma tissues and its effect on the migration and invasion of glioma cells.Methods:The expression of RAC3 in 57 glioma patients and their adjacent tissues from the First People’s Hospital of Shangqiu was detected by immunohistochemical assay. According to the experimental requirements, brain glioma cells U87MG were divided into experimental group and control group. The experimental group U87MG cells were transfected with RAC3-siRNA plasmid, and the control group U87MG cells were transfected with MOCK-siRNA plasmid. RAC3 mRNA in each group was detected by fluorescence quantitative PCR. The expressions of RAC3 and MMP2 in each group were detected by Western blot. Transwell was used to detect the migration and invasion ability of cells in each group.Results:The positive rate of RAC3 in glioma patients was 89.47% (51/57 cases) , and the expression rate in paracancer tissues was 14.04% (8/57 cases) . The expression rate of RAC3 in glioma tissues was significantly higher than that in paracancer tissues, with statistical significance ( P<0.01) . After siRNA transfection, mRNA expression of RAC3 in experimental group and control group was 1.23±0.20 and 0.43±0.12, and protein expression of RAC3 was 1.19±0.11 and 0.23±0.08, respectively. The expression of MMP2 protein was 1.19±0.11 and 0.23±0.08, respectively. The expression of MMP2 in experimental group was significantly decreased ( P<0.05) . Transwell assay showed that the number of invasive cells in experimental group and control group U87MG cells was (22±5) and (45±8) , and the number of migratory cells was (34±6) and (90±11) , respectively. In experimental group, U87MG cell migration and invasion ability decreased significantly (both P<0.05) . Conclusion:The high expression of RAC3 in glioma tissues may be related to the malignant degree of development, and affect the migration and invasion ability of glioma cells by regulating the expression of MMP2.
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Objective @#To explore the possible mechanism of glycyrrhetinic acid on inhibiting malignant biological behaviors of melanoma by Wnt / β-catenin pathways．@*Methods@#The melanoma cells B16-F10 were selected as the research objects．The concentration gradient tests (0，1，2，4 μmol /L) were conducted by MTT．The cells given cisplatin intervention was enrolled as positive controls．The cells invasion and migration were detected by Transwell chamber assay．The expression levels of Wnt / β-catenin pathway proteins，invasion and migration related proteins (MMP-2，MMP-9) in B16-F10 cells were detected by Wester blot．The xenograft models of nude mice were constructed，and they were divided into control group (without drugs treatment) and glycyrrhetinic acid group (40 mg / kg) ．The growth of tumor tissues，and expression levels of Wnt / β-catenin pathway proteins，invasion and migration related proteins were observed. @*Results @#MTT results showed that glycyrrhetic acid could inhibit the proliferation of B16-F10 cells in a concentration-dependent manner．The inhibition effect of glycyrrhetic acid ( ≥2 μmol /L) was significant on the proliferation of B16-F10 cells (P ＜0. 05) ．The results of Transwell chamber assay showed that compared with control group，invasion and migration abilities of B16-F10 cells were significantly reduced after treatment with glycyrrhetinic acid (2，4 μmol /L) (P ＜0. 05) ．Wester blot results showed that compared with those without glycyrrhetinic acid treatment，expression levels of MMP-9 ，MMP-2，Wnt1 and β-catenin protein in B16- F10 cells significantly decreased after treatment with glycyrrhetinic acid (2，4 μmol /L) (P＜0. 05) ．The results of tumor-bearing assay showed that compared with control group ，weight and volume of tumors significantly decreased in glycyrrhetinic acid group，and expression levels of Wnt1 ，β-catenin，MMP-9 and MMP-2 proteins also significantly decreased (P＜0. 05) ．@*Conclusion @#Glycyrrhetinic acid can significantly inhibit the malignant biological behaviors of melanoma in vitro and vivo．And its mechanism may be related to inhibiting the activation of Wnt / β-catenin signaling pathways.
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AIM:To study the effect of Aurora protein kinase inhibitor VX-680 on homogeneous adhesion and migration ability in human hepatocellular carcinoma cell line HepG 2.METHODS:The HepG2 cell were divided into ex-perimental group and control group, respectively.VX-680 was used in experimental groups at 3 concentrations(3.125 μmol/L group,6.25 μmol/L group and 12.5 μmol/L group).DMSO was used in the control group.The effects of VX-680 at different concentrations on the adhesion ability of human hepatocellular carcinoma HepG 2 cells were observed by cell slow aggregation test and separation experiment.The effects of VX-680 at different concentrations on the migration ability of HepG2 cells was detected by wound healing assay.The expression of E-cadherin in HepG2 cells was detected by Western blot.RESULTS:The results of the slow aggregation test showed that compared with the control group,the number of cell clumps formed in experimental groups was significantly decreased(P<0.01).The results of separation experiment showed that the ratio of NTC/NTEgradually decreased with the increased concentration of VX-680.The results of wound healing as-say showed that as the concentration of VX-680 increased, the cell scratch healing ability gradually weakened compared with control group.The results of Western blot showed that the protein expression of E-cadherin in the HepG2 cells in-creased with the increased concentration of VX-680(P<0.05).CONCLUSION:VX-680 increases the homogeneous ad-hesion and inhibits the migration of HepG 2 cells.
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Objective To investigate the effects of semaphorin 4D(Sema4D) on the proliferation,migration and angiogenic of human pancreatic carcinoma cells.Methods Sema4D-siRNA was designed and synthesized and transfected into human pancreatic carcinoma cells.After 48 hours of transient infection,the changes of expression of Sema4D mRNA before and after transfection were detected by reverse transcription-polymeruse chain reaction method.And after 72 hours of transient infection,the changes of expression of Sema4D protein before and after transfection were detected by Western blot method.The changes of growth of the transfected cells were observed by methyl thiazolyl terazolium assay.Using transwell migration test and scratch repair test to detect the changes of migration ability of human pancreatic carcinoma cells after transfection.Using tubule formation assay to observe the effect of supernatant of pancreatic carcinoma cell cultures on angiogenesis after transfection.Results Compared with the negative control group and blank control group,the expression of Sema4D mRNA and Sema4D protein and the growth rate of pancreatic carcinoma cells decreased significantly (P ＜ 0.05).In transwell migration test and scratch repair test,it was observed that Pancreatic cancer cells penetrating cell number and scratch repair rate were significantly lower than that in negative control group and blank control group (P ＜ 0.05).Tubule formation assay showed that there were significant differences in angiogenesis numbers among siRNA transfection group(0.5 ± 0.02),negative control group(1.45 ± 0.60) and blank control group (1.37 ± 0.52) (P ＜ 0.05).Conclusion Sema4D-siRNA can induce RNA interference in pancreatic carcinoma cells and down-regulate the expression of Sema4D gene,which can inhibit the proliferation of pancreatic carcinoma cells,significantly reduce the migration ability of pancreatic carcinoma ceils and inhibit angiogenesis.