同源异型盒1 (sine oculis homeobox homolog 1,SIX1)基因参与多种肿瘤调控,在乳腺癌发生发展过程中发挥了重要作用,但具体机制仍有待探讨.本研究目的 在于分析SIX1基因在乳腺癌中的表达情况,研究其在乳腺癌细胞增殖与侵袭过程中所发挥的作用并探讨作用的机制.TCGA数据库和GEPIA2在线数据库分析发现,SIX1在乳腺癌中的表达高于正常组织,且在乳腺癌的不同分子亚型Basal-like、HER2、Luminal A、Luminal B中也证实了这一点(P＜0.05);HPA数据库分析发现,SIX1的免疫组化蛋白质表达水平高于正常乳腺组织,SIX1基因在乳腺癌中高表达,差异具有统计学意义(P＜0.001).在乳腺癌细胞株MDA-MB-231中,干扰SIX1的表达及在MCF-7中过表达SIX1后,生长曲线及EdU实验结果显示:敲低SIX1后乳腺癌细胞的生长增殖受到抑制,而过表达SIX1则会发挥促进作用(生长曲线实验:P＜0.05;EdU实验:P＜0.001);Transwell结果表明,SIX1能够增强乳腺癌细胞的侵袭能力(P＜0.001).利用TCGA数据库,根据SIX1基因表达值的上下四分位定义SIX1基因高表达及低表达人群,对两组队列进行差异基因分析,发现差异基因与代谢、干细胞调控等通路相关;对敲低SIX1的乳腺癌细胞系进行转录物组测序(RNA-sequencing,RNA-seq),并对测序结果进行一系列生物信息学分析发现,SIX1与代谢、干细胞调控以及EMT等通路密切相关.挑选代表性基因MYC、SNAI2、EGFR,通过已发表的GEO临床数据集以及KM-plot-ter验证靶基因与SIX1表达的相关性及与临床乳腺癌病人的预后关系,我们发现:SIX1与MYC、SNAI2、EGFR表达正相关.高表达的SIX1、MYC、SNAI2与EGFR不利于乳腺癌病人的生存.利用GCBI、GeneMANIA及String在线工具,我们发现了SIX1基因相互作用的蛋白质和相关的lncRNA及miRNA.综上,本研究中初步揭示了SIX1在乳腺癌中发挥的作用及其调控机制,为进一步深入研究该基因提供了线索.

乳腺导管原位癌是一种异质性肿瘤,按其形态学特点、免疫组织化学染色、分子标志物以及临床表现来看,虽然肿瘤的生物学行为各不相同,但共同特征是病变主要局限在乳腺导管-小叶系统内[1].乳腺导管-小叶系统被覆双层细胞结构:内层的腺腔上皮细胞和外层(基底面)的肌上皮细胞,一旦腺腔来源的癌细胞突破外层(基底面)的肌上皮细胞,就会发展为浸润性癌.故有不少学者把导管原位癌看作是浸润性癌的癌前病变[2].

Objective To evaluate the correlation of skin prick test (SPT)of dust mites with serum specific IgE(sIgE) and age.Methods A total of 160 patients with allergic rhinitis received SPT.And sIgE was detected.The correlation of skin prick test (SPT)of dust mites with serum specific IgE(sIgE)and age was analyzed.Results There was posi-tive correlation between the results of SPT and grades of sIgE and negative correlation between the results of SPT and age.Conclusion There was significant correlation between SPT of dust mites and sIgE and age.

Objective:To analyze the effect of folate deficiency on protein expression of embryonic rat neural stem cells(NSCs).Methods:Embryonic rat NSCs were cultured in serum-free suspension culture and were identified.Individual cultures were assigned to one of 3 treatment groups according to the concentration of folate in the medium:control group,folate deficiency group(Folate-D) and folate supplementation group(Folate-L).The final folate concentrations were 4,0.6 and 8 mg/L respectively.Protein expressions were detected by two-dimensional electrophoresis(2-DE) in 3 groups.Results:The stable and repeatable 2-DE maps of NSCs were successfully established.Compared with control group,2 spots of proteins were lowly expressed,4 spot of proteins were highly expressed in Folate-D group.And 2 spots of proteins were lowly expressed,1 spot of proteins was highly expressed in Folate-L group.Conclusion:Folate deficiency or supplement may impact NSCs protein expression.The isolation of differentially expressed proteins is useful for exploring the mechanism of folate on proliferation and differentiation of NSCs.

Objective To explore the role of metastasis associated protein-1(Mta-1)and vascular endothelial growth factor(VEGF) expression in the occurrence and development of osteosarcoma after chemotherapy.And to investigate the relation between MTA1 and VEGF combinative expression and the prognosis in osteosarcoma patients after chemotherapy.Methods SP immunohistochemical technique was used to detect the expression of MTA1 and VEGF in the residual tumor cells from 31 osteosarcoma patients after chemotherapy.Results There were statistical significance of MTA1 and VEGF combinative expression for the prognosis in osteosarcoma patients after chemotherapy.Conclusion Both negative of MTA1 and VEGF are better for the prognosis in osteosarcoma patients after chemotherapy.While positive expression for both of them means poor prognosis.The results indicate that Mta-1 and VEGF combinative expression could be used to assess the prognosis of patients with osteosarcoma after chemotherapy.

BACKGROUND:Previous studies have verified that under normal culture of neural stem cells (NSCs),folic acid can accelerate proliferation of NSCs by phosphorylation of mitogen activated protein kinase path activation ERK1/2.OBJECTIVE:To investigate the effect of folic acid on NSC extracellular signal regulatory protein kinase pERK1/2 under hypoxic condition.METHODS:NSCs from Neonatal rats were cultured in vitro by serum-free culture method,and incubated in a flask at 1×108/L.Except normal control group,self-made hypoxia equipment was used in the hypoxia model,folic acid deficiency and folic acid supplemented groups at day 3.At 37 ℃,hypoxia culture was conducted in the thermostat for 6 hours.The contents of folic acid were 4 mg/L,4 mg/L,0.65 mg/L,8 mg/L in the four groups.Cells following 6 days were collected to count the density using trypan blue.RT-PCR was utilized to detect pERK1/2 mRNA expression.Western blot assay was employed to determine pERK1/2 protein expression.RESULTS AND CONCLUSION:Compared with the normal control group,the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were decreased significantly in the hypoxia model group.Compared with the hypoxia model group,the proliferation of NSCs and the expression of ERK1/2mRNA and pERK1/2 protein were increased in the folic acid supplemented group,whereas decreased in the folic acid deficiency group.There were significant differences among groups (P < 0.001).Above-described results verified that folic acid supplementation can activate ERK1/2 phosphorylatin and accelerate proliferation of NSCs under hypoxia condition.

Objective To study the effects of folate(Fol) on proliferation,apoptosis and the antioxidative capacity of the NSCs under hypoxia in vitro.Method The NSCs taken from infant rats brain were cultured by serum-free medium in vitro and divided into four group which were normal control group(NC),hypoxia group(Hyp),Hpy+Fol-D(folate deficient)group and Hpy+Fol group.Except NC group,the other groups were cultured in hypoxia condition for 6 h.The proliferation of NSCs after hypoxia damage was detected by MTT test.The NSCs were collected after being cultured 6 d and then the activity of SOD and GSH-PX was examined.The expression of activated caspase-3 was detected by Western blot.Results Compared with NC group,the proliferation of NSCs and the activity of SOD and GSH-PX decreased significantly after hypoxia,while the expression of activated caspase-3 increased0 The proliferation of NSCs and the activity of SOD and GSH-PX in Hyp group were significantly higher than Hyp+Fol-D group but much lower than Hpy+Fol group(P0.05),while the expression of activated caspase-3 was much higher than Hpy+Fol group but much lower than Hpy+Fol-D group(P0.05).Conclusion Folate deficiency can aggravate the damage of NSCs injured after hypoxia,and folate supplement can promote the proliferation of NSCs,improve the antioxidative capacity and inhibit the apoptosis of NSCs induced by hypoxia,so that folate can promote the recovery of hypoxia damaged NSCs in vitro.

Objective:To explore the effect of folic acid (FA) on the mRNA and protein expression level of Hes5 of neural stem cells (NSCs) from neonatal rats in vitro after hypoxia damage.Methods:Cultured NSCs were randomly divided into four groups:normal control group,hypoxia model group,hypoxia FA-supplemented group and hypoxia FA deficiency group.On the third day of proliferation,the cells of three groups excepting the normal control group were cultured in hypoxia condition for 6 hours,then continued to culture until the sixth day when the NSCs of the groups were collected.The cell density was calculated by Trypan blue,the expression of mRNA was detected by RT-PCR and the expression of protein was measured by Western blot.Results:Trypan blue staining illustrated the cell density in hypoxia FAsupplemented group was higher than in other three groups,hypoxia FA deficiency group was the lowest among the four groups,the differences between them were significant (P0.05).The results of RT-PCR showed that the mRNA expression of hypoxia FA-supplemented group was more than other groups,that of hypoxia FA deficiency group was the minimal of all groups,the differences between them were significant (P 0.05).The consequences of Western blot illustrated that the protein expression of hypoxia FA supplemented group was the most of all four groups while hypoxia FA deficiency group was the least amid all groups,the differences between them were significant (P 0.05).Conclusion:The hypoxia model of NSCs in vitro is established successfully,folic acid may stimulate proliferation of hypoxia damaged NSCs efficiently,which could supply the evidence for precaution and therapy of hypoxic ischemic damage.