ABSTRACT
Objective Non-muscle myosin heavy chain 9 (MYH9) plays an important regulatory role in the development of tumor.This study aimed to explore the expression of MYH9 in osteosarcoma tissues and its effects on epithelial-mesenchymal transition and invasion of osteosarcoma cells.Methods We collected 52 cases of osteosarcoma tissues and para-carcinoma tissues at 5 cm form the edge of the tumor.RT-PCR and immunohistochemistry were used to analyze the expression level of MYH9 mRNA and protein in the osteosarcoma tissues and para-carcinoma tissues.MYH9 shRNA plasmid was transfected into U2-OS cells to silenced the expression of MYH9, after transfected, the cells were divided into three groups: the normal U2-OS cells were the control group, the U2-OS cells transfected with empty plasmid were the empty group and U2-OS cells transfected with MYH9 shRNA were interference group.RT-PCR was used to detect the changes of MYH9 mRNA levels in the U2-OS cells, the protein level of MYH9, EMT related protein E-cadherin and Vimentin were detected by Western blot, and the ability of cell invasion was evaluated by Transwell assay.Results The results of RT-PCR showed that the relative expression MYH9 mRNA in para-carcinoma tissues(1.526±0.148) was significantly lower than that in cancer tissues (3.547±0.195) (P<0.05).The results of immunohistochemistry showed that MYH9 protein was mainly expressed in cytoplasm, and the expression in cancer tissues was significantly higher than that in para-carcinoma tissues, the positive expression rate were 59.6%(31/52) and 26.9%(14/52) respectively, the difference was statistically significant(P<0.05).The results of Western blot showed that the relative expression of MYH9 mRNA in interference group was significantly lower than that in control group and empty group (P<0.05) after silenced MYH9 gene, and compared with the control group, the E-cadherin in U2-OS cells was significantly up-regulated but the Vimentin was down-regulated.After 48h, all of the groups had cells through the microfiltration membrane, the numbers of cells through the microfiltration membrane in interference group(41.2±15.1) was significantly lower than that in control group(117.3±12.4) and empty group(193.5±14.7) (P<0.05).Conclusion The expression of MYH9 protein in osteosarcoma tissues was significantly higher than that in para-carcinoma t tissues, silenced MYH9 gene can reduce the invasive ability of osteosarcoma by reducing the epithelial interstitial transition.
ABSTRACT
Objective To investigate thecorrelation between nonmusle myosin heavy chain 9 gene (MYH9) rs12107,signal transducer and activator of transcription (STAT4) rs3024912, Urokinase plasminogen activator (uPA) rs4065 single nucleotide polymorphism and idiopathic Uighur membranous nephropathy (IMN). Methods Patients admittedby People′s Hospital of Xinjiang Uyghur Autonomous Region from June 2011 to May 2015 were selected in the research,of which 45 with IMN (group A),45 patients with IgA nephropathy (group B) and 45 healthy controls(group C). The polymorphisms of rs12107,rs3024912 and rs4065 were measured with direct sequencing, in order to analyzing the correlation between genotype and allele with IMN. Results Group Ars12107 (MYH9) locus genotype CC, C allele (48.9%, 65.6%) frequency were higher than those in group B (13.3%, 33.3%) and group C (20.0%, 46.7%), and the difference was statistically significance (P 0.05). rs3024912 GG genotype patients showed higher risk of renal failure compared with non-GG genotype patients (95% CI:1.48-26.83, P = 0.013). Only TT genotype was detected on rs4065 locus. TC and CC genotype were not detected. Conclusions MYH9 gene rs12107 locus CC genotype and C allele are associated with susceptibility to IMN in Xinjiang Uygur, and CC genotypes associated with renal function. rs3024912 (STAT4)GG genotype are not susceptibility gene,but associated with renal function in patients with IMN.
ABSTRACT
Objective To study the association of MYH9 gene single nucleotide polymorphism (SNP) with clinical manifestation,pathology and prognosis of IgA nephropathy (IgAN) patients of Han nationality population in Inner Mongolia Autonomous Region.Method One hundred and forty-eight IgAN patients proven by biopsy were enrolled in the study.Fifty-six patients were followed up for 1-97 months.DNA was extracted from the peripheral blood of above patients.PCR restriction fragment length polymorphism (RFLP) assay was used to detect the single nucleotide polymorphisms of MYH9 gene Rs3752462,Rs4821480 sites.Association of different genotypes with clinical features,pathology and prognosis im patients with IgA nephropathy was examined.Result (1) Rs3752462 site was consistent with Hardy-Weinberg equilibrium,while Rs4821480 site did not meet the Hardy-Weinberg equilibrium.(2) IgAN patients with MYH9 gene Rs3752462 site TF genotype had lower systolic blood pressure as compared to those with CC +CT genotype (P＜0.05).There were significant differences in systolic blood pressure,diastolic blood pressure and age between patients with Rs4821480 site GG genotype and patients with TT or GT genotype (P＜0.05).There were no significant differences in Scr,Ccr,plasma albumin,hemoglobin,microscopic hematuria,proteinuria,pathological HASS classification,pathological lesion among Rs4821480 site GG,TT,GT genotypes.(3) Kaplan-Meier survival analysis revealed the time from renal biopsy to renal function decline was shorted in patients with Rs3752462 site CC genotype and Rs4821480 site TT genotype.Conclusions C allele of MYH9 gene Rs3752462 site is an independent risk factor of high blood pressure damage in IgAN patients.Polymorphism of 3 genotypes of MYH9 gene Rs4821480 site is associated to the prognosis of patients.Carrying Rs3752462 site C allele and Rs4821480 site T allele may affect the prognosis of patients.