PRDM4通过PI3K/AKT信号通路抑制宫颈癌细胞的侵袭和转移
|
摘要
目的研究PR-domain蛋白转录因子家族成员PRDM4在宫颈癌发生发展中的作用和临床意义。方法采用免疫组化法检测30例正常宫颈组织和30例宫颈癌组织中PRDM4的表达。在宫颈癌细胞系Si Ha和He La中外源性过表达和干扰PRDM4,采用Transwell小室实验检测其对宫颈癌细胞Si Ha和He La侵袭转移能力的影响。采用转录本测序结合基因富集方法分析PRDM4过表达Si Ha细胞和对照细胞的差异信号通路和基因表达,并通过real-time PCR进行验证。PI3K/AKT激动剂胰岛素样生长因子1(insulin-like growth factor 1,IGF1)处理PRDM4过表达的Si Ha和He La细胞后,检测细胞的侵袭转移能力。结果 PRDM4在宫颈癌组织中的表达显著低于正常宫颈组织,并且与宫颈癌患者的临床分期和淋巴结转移呈负相关(P<0.05);外源性的过表达/干扰PRDM4能够显著抑制宫颈癌细胞Si Ha和增强He La的体外侵袭和转移能力。转录本测序结合基因富集结果显示PRDM4过表达Si Ha细胞中PI3K/AKT信号通路及下游基因的表达受到抑制。利用PI3K/AKT激动剂显著促进了PRDM4过表达所导致的细胞侵袭和转移抑制作用。结论 PRDM4在宫颈癌中的表达下调,与患者的临床分期及预后相关,机制研究显示PRDM4通过抑制PI3K/AKT信号通路抑制宫颈癌细胞的侵袭转移能力。
Objective To explore the role and clinical significance of the PR-domain transcription factor PRDM4 in the development of cervical cancer.Methods Immunohistochemical assay was used to detect the expression of PRDM4 in 30 cases of normal cervical tissues and 30 cases of cervical carcinoma.Cell invasion and metastasis abilities were detected by Transwell assay in PRDM4 over-expression or interference cervical cancer cells Si Ha and He La and the controls respectively.The RNA sequencing and gene enrichment were used to analyze the differential signaling pathway and gene expression and then verified by real-time PCR.PRDM4 over-expressed Si Ha and He La cells were treated by PI3K/AKT agonists insulin-like growth factor 1(IGF1).Results The expression of PRDM4 in cervical cancer tissues was significantly lower than that in normal tissues,and negatively correlated with the clinical stage and lymph node metastasis(P < 0.05).The cell invasion and metastasis abilities of Si Ha cells were significantly inhibited after PRDM4 overexpression and enhanced after interference,respectively.The RNA sequencing results showed that PRDM4 inhibited the activation of PI3K/AKT signaling pathway and downstream genes expression.PI3K/AKT agonists significantly promoted the inhibition of invasion and metastasis by PRDM4 overexpression.Conclusion This study suggests that the expression of PRDM4 is down-regulated and negatively related with clinical stage and prognosis of cervical cancer.PRDM4 may inhibit the cell invasion and metastasis by inhibiting the PI3K/AKT signaling pathway.
Objective To explore the role and clinical significance of the PR-domain transcription factor PRDM4 in the development of cervical cancer.Methods Immunohistochemical assay was used to detect the expression of PRDM4 in 30 cases of normal cervical tissues and 30 cases of cervical carcinoma.Cell invasion and metastasis abilities were detected by Transwell assay in PRDM4 over-expression or interference cervical cancer cells Si Ha and He La and the controls respectively.The RNA sequencing and gene enrichment were used to analyze the differential signaling pathway and gene expression and then verified by real-time PCR.PRDM4 over-expressed Si Ha and He La cells were treated by PI3K/AKT agonists insulin-like growth factor 1(IGF1).Results The expression of PRDM4 in cervical cancer tissues was significantly lower than that in normal tissues,and negatively correlated with the clinical stage and lymph node metastasis(P < 0.05).The cell invasion and metastasis abilities of Si Ha cells were significantly inhibited after PRDM4 overexpression and enhanced after interference,respectively.The RNA sequencing results showed that PRDM4 inhibited the activation of PI3K/AKT signaling pathway and downstream genes expression.PI3K/AKT agonists significantly promoted the inhibition of invasion and metastasis by PRDM4 overexpression.Conclusion This study suggests that the expression of PRDM4 is down-regulated and negatively related with clinical stage and prognosis of cervical cancer.PRDM4 may inhibit the cell invasion and metastasis by inhibiting the PI3K/AKT signaling pathway.
引文
[1]Ferlay J,Soerjomataram I,Dikshit R,et al.Cancer incidence and mortality worldwide:sources,methods and major patterns in GLOBOCAN 2012[J].Int J Cancer,2015,136(5):E359-386.
[2]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[3]Bogani D,Morgan MA,Nelson AC,et al.The PR/SET domain zinc finger protein Prdm4 regulates gene expression in embryonic stem cells but plays a nonessential role in the developing mouse embryo[J].Mol Cell Biol,2013,33(19):3936-3950.
[4]Briknarova K,Atwater DZ,Glicken JM,et al.The PR/SET domain in PRDM4 is preceded by a zinc knuckle[J].Proteins,2011,79(7):2341-2345.
[5]Song NJ,Choi S,Rajbhandari P,et al.Prdm4 induction by the small molecule butein promotes white adipose tissue browning[J].Nat Chem Biol,2016,12(7):479-481.
[6]Tunbak H,Georgiou C,Guan C,et al.Zinc fingers 1,2,5 and6 of transcriptional regulator,PRDM4,are required for its nuclear localisation[J].Biochem Biophys Res Commun,2016,474(2):388-394.
[7]Giguere V,Yang N,Segui P,et al.Identification of a new class of steroid hormone receptors[J].Nature,1988,331(6151):91-94.
[8]Yan Z,Xiong Y,Xu W,et al.Identification of recurrence-related genes by integrating microRNA and gene expression profiling of gastric cancer[J].Int J Oncol,2012,41(6):2166-2174.
[9]Yang XH,Huang S.PFM1(PRDM4),a new member of the PR-domain family,maps to a tumor suppressor locus on human chromosome 12q23-q24.1[J].Genomics,1999,61(3):319-325.
[10]Oyervides-Mu1oz MA,Pérez-Maya AA,Rodríguez-Gutiérrez HF,et al.Understanding the HPV integration and its progression to cervical cancer[J].Infect Genet Evol,2018,7(61):134-144.
[11]Mzoughi S,Tan YX,Low D,et al.The role of PRDMs in cancer:one family,two sides[J].Curr Opin Genet Dev,2016,2(36):83-91.
[12]Liu H,Dai XM,Cao XL,et al.PRDM4 mediates YAP-induced cell invasion by activating leukocyte-specific integrinβ2 expression[J].EMBO Reports,2018,5(19):e45180.
[13]Honardoost M,Rad SMAH.Triangle of AKT2,miRNA,and Tumorigenesis in Different Cancers[J].Appl Biochem Biotechnol,2018,185(2):524-540.
[2]Chen W,Zheng R,Baade PD,et al.Cancer statistics in China,2015[J].CA Cancer J Clin,2016,66(2):115-132.
[3]Bogani D,Morgan MA,Nelson AC,et al.The PR/SET domain zinc finger protein Prdm4 regulates gene expression in embryonic stem cells but plays a nonessential role in the developing mouse embryo[J].Mol Cell Biol,2013,33(19):3936-3950.
[4]Briknarova K,Atwater DZ,Glicken JM,et al.The PR/SET domain in PRDM4 is preceded by a zinc knuckle[J].Proteins,2011,79(7):2341-2345.
[5]Song NJ,Choi S,Rajbhandari P,et al.Prdm4 induction by the small molecule butein promotes white adipose tissue browning[J].Nat Chem Biol,2016,12(7):479-481.
[6]Tunbak H,Georgiou C,Guan C,et al.Zinc fingers 1,2,5 and6 of transcriptional regulator,PRDM4,are required for its nuclear localisation[J].Biochem Biophys Res Commun,2016,474(2):388-394.
[7]Giguere V,Yang N,Segui P,et al.Identification of a new class of steroid hormone receptors[J].Nature,1988,331(6151):91-94.
[8]Yan Z,Xiong Y,Xu W,et al.Identification of recurrence-related genes by integrating microRNA and gene expression profiling of gastric cancer[J].Int J Oncol,2012,41(6):2166-2174.
[9]Yang XH,Huang S.PFM1(PRDM4),a new member of the PR-domain family,maps to a tumor suppressor locus on human chromosome 12q23-q24.1[J].Genomics,1999,61(3):319-325.
[10]Oyervides-Mu1oz MA,Pérez-Maya AA,Rodríguez-Gutiérrez HF,et al.Understanding the HPV integration and its progression to cervical cancer[J].Infect Genet Evol,2018,7(61):134-144.
[11]Mzoughi S,Tan YX,Low D,et al.The role of PRDMs in cancer:one family,two sides[J].Curr Opin Genet Dev,2016,2(36):83-91.
[12]Liu H,Dai XM,Cao XL,et al.PRDM4 mediates YAP-induced cell invasion by activating leukocyte-specific integrinβ2 expression[J].EMBO Reports,2018,5(19):e45180.
[13]Honardoost M,Rad SMAH.Triangle of AKT2,miRNA,and Tumorigenesis in Different Cancers[J].Appl Biochem Biotechnol,2018,185(2):524-540.