宫颈癌组织DcR3的表达与高危型人乳头状瘤病毒感染的相关性研究

目的  观察宫颈癌组织诱捕受体3（decoy receptor 3，DcR3）表达与高危型人乳头状瘤病毒（high-risk human papilloma virus，HR-HPV）感染的相关性。 方法  分别采用免疫组织化学（streptavidin-perosidase，SP）法和第2代杂交捕获法检测35例正常宫颈组织（normal cervical tissues，NCE）、39例宫颈上皮内瘤变（cervical intraepithelial neoplasm，CIN）及44例宫颈癌组织（cervical squamous epithelial carcinoma，CSES）DcR3和HR-HPV的表达。设计并合成靶向HR-HPV16 E7基因的HR-HPV16 E7小干扰RNA（siRNA）和阴性对照HR-HPV16 E7 siRNA，脂质体法转染SiHa细胞，实时荧光定量聚合酶链式反应（real-time polymerase chain reaction，RT-PCR）法和蛋白质印迹（western blot，WB）法检测HR-HPV16 E7基因沉默对DcR3 mRNA和蛋白表达的影响，四甲基偶氮唑盐比色（methylthiazolyldiphenyl-tetrazolium bromide，MTT）法检测细胞增殖情况。 结果  NCE、CIN及CSES组DcR3蛋白阳性表达率分别为8.6%（3/35）、48.7%（19/39）、77.3%（34/44），表达呈逐渐增高趋势，差异具有统计学意义（χ²=36.942，P<0.001）。NCE、CIN及CSES组HR-HPV阳性检出率分别为5.7%（2/35）、56.4%（22/39）、93.2%（41/44），差异具有统计学意义（χ²=60.322，P<0.001）。DcR3表达与HR-HPV感染呈正相关（r=0.893，P=0.004）。HR-HPV基因沉默后DcR3在mRNA和蛋白水平的表达均明显降低，且细胞增殖受到抑制。 结论  DcR3在宫颈癌中异常高表达，其表达升高与HR-HPV感染密切相关，提示DcR3可能参与了宫颈癌的发生发展。siRNA技术成功诱导HR-HPV基因沉默可通过下调DcR3的表达抑制人宫颈癌SiHa细胞增殖。

Relationship between the expression of DcR3 and high-risk human papilloma virus infection in cervical carcinoma

  Objective  To explore the relationship between the expression of decoy receptor 3 (DcR3) and high-risk human papilloma virus(HR-HPV) infection in cervical carcinoma.  Methods  Immunohistochemistry and hybird capture Ⅱ assay were used to detect the expression of DcR3 and HR-HPV in 35 cases of normal cervical tissues(NCE), 39 cases of cervical intraepithelial neoplasm(CIN) and 44 cases of cervical squamous epithelial carcinoma(CSES). Specific HR-HPV16 E7 siRNA and nonspecific HR-HPV16 E7 siRNA were synthesized and transfected to SiHa cells by Lipofectamine. The expression of DcR3 at mRNA and protein levels was examined by real-time polymerase chain reaction and western blot. The growth inhibition was examined by MTT assay.  Results  In NCE, CIN and CSES, the positive expression rates of DcR3 were 8.6%(3/35), 48.7%(19/39) and 77.3%(34/44), respectively, and the expression intensity was increasing(χ2=36.942, P<0.001). In NCE, CIN and CSES, the infection rates of HR-HPV were 5.7%(2/35), 56.4%(22/39) and 93.2%(41/44), respectively(χ2=60.322, P<0.001). The protein expression of DcR3 was positively correlated with the infection of HR-HPV in CSES(r=0.893, P=0.004).  Conclusions  DcR3 was highly expressed in cervical cancer. Its expression was positively correlated with HR-HPV infection, which may contribute to the occurrence and development of cervical cancer. HR-HPV silencing inhibited cellular growth and proliferation by down-regulating the expression of DcR3.