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

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

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-HPV16E7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(X^2=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(X^2=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Si Ha细胞增殖。

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 Si Ha 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( χ^2= 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( χ^2= 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.