血浆M-CSF、MMP9、TIMP-1水平及HPV阳性大鼠宫颈癌增殖能力的关系研究  

Study on the Relationship between Plasma Levels of M-CSF,MMP9,TIMP-1 and Cervical Cancer Proliferation in HPV-positive Rats

作　　者：许林波 梁丽斯 王蓓[3] 邓晓红 白昌民 XU Lin-bo;LIANG Li-si;WANG Bei;DENG Xiao-hong;BAI Chang-min(Medical Laboratory Center,Northwest Women's and Children's Hospital,Xi'an,Shaanxi,710000,China;Department of Laboratory Medicine,Xi'an Ninth Hospital,Xi'an,Shaanxi,710014,China;Central Laboratory of Xi'an Children's Hospital,Xi'an,Shaanxi,710000,China;Department of Gynecology,Northwest Women and Children's Hospital,Xi'an,Shaanxi,710000,China)

机构地区：[1]西北妇女儿童医院医学检验中心,陕西西安710000 [2]西安市第九医院检验科,陕西西安710014 [3]西安市儿童医院中心实验室,陕西西安710000 [4]西北妇女儿童医院妇科,陕西西安710000

出　　处：《现代生物医学进展》2022年第8期1439-1443,共5页Progress in Modern Biomedicine

基　　金：陕西省重点研发计划项目(2018SF-068)。

摘　　要：目的:探究血浆巨噬细胞集落刺激因子(Macrophage colony stimulating factor,M-CSF)、基质金属蛋白酶9(Matrix metalloproteinase 9,MMP9)及其组织抑制因子1(tissue inhibitor of the metalloproteinases,TIMP1)水平及人乳头瘤病毒(Human papilloma virus,HPV)阳性大鼠宫颈癌增殖能力的关系。方法:20只健康雌性Wistar白化大鼠根据实验目的分为两组:对照组(异种移植时注射Si Ha细胞作为对照实验,n=10)和观察组(将转染sh-M-CSF、sh-MMP9和sh-TIMP-1的Si Ha细胞注射大鼠的子宫颈,n=10)。通过ELISA测定大鼠血浆M-CSF、MMP9和TIMP-1的水平。通过PCR检测实验大鼠中M-CSF、MMP9和TIMP-1的m RNA表达。使用数字游标卡尺分析大异种移植大鼠肿瘤体积生长。第3、4、5周分别处死并切除大鼠肿瘤进行称重。通过免疫组织化学分析肿瘤组织中增殖细胞核抗原(Proliferating cell nuclear antigen,PCNA)、pAKT和pSTAT3的蛋白表达。通过免疫组织化学染色和TUNEL染色分别确定Ki67阳性细胞数量及凋亡细胞数量。结果:观察组较对照组M-CSF、MMP9和TIMP-1的水平降低(P<0.05)。观察组较对照组M-CSF、MMP9和TIMP-1的m RNA表达降低(P<0.05)。随着时间的增加,两组大鼠肿瘤体积均增加。1周和2周对照组和观察组大鼠肿瘤体积比较无差异(P>0.05),第3周、第4周和第5周,观察组较对照组大鼠肿瘤体积降低(P<0.05)。观察组较对照组大鼠体内肿瘤重量减少(P<0.05)。观察组较对照组PCNA、pAKT和pSTAT3的蛋白表达量降低(P<0.05)。观察组较对照组Ki67阳性细胞数量降低,凋亡细胞升高(P<0.05)。结论:降低血浆M-CSF、MMP2和TIMP1水平可促进HPV阳性大鼠宫颈癌细胞凋亡,有效抑制细胞增殖。Objective:To explore the relationship between the levels of plasma M-CSF,MMP9,TIMP-1 and the proliferation of cervical cancer in HPV-positive rats.Methods:Twenty healthy female Wistar albino rats were divided into two groups according to the purpose of the experiment:the control group(injecting Si Ha cells during xenotransplantation as a control experiment,n=10)and the observation group(the Si Ha cells transfected with sh-M-CSF,sh-MMP9 and sh-TIMP-1)Inject into the cervix of rats,n=10).The plasma levels of M-CSF,MMP9 and TIMP-1 were determined by ELISA.The m RNA expression of M-CSF,MMP9 and TIMP-1 in experimental rats was detected by PCR.The digital vernier caliper was used to analyze the tumor volume growth of large xenograft rats.At the 3rd,4th,and 5th week,the rats were sacrificed and the tumors were excised and weighed.The protein expression of Proliferating cell nuclear antigen(PCNA),pAKT and pSTAT3 in tumor tissues was analyzed by immunohistochemistry.The number of Ki67-positive cells and the number of apoptotic cells were determined by immunohistochemical staining and TUNEL staining,respectively.Results:Compared with the control group,the observation group had lower levels of M-CSF,MMP9 and TIMP-1(P<0.05).The m RNA expression of M-CSF,MMP9 and TIMP-1 in the observation group was lower than that in the control group(P<0.05).With the increase of time,the tumor volume of the two groups of rats increased.There was no difference in tumor volume between the control group and the observation group at 1 week and 2 weeks(P>0.05).At the 3rd,4th and 5th week,the observation group had a lower tumor volume than the control group(P<0.05).The observation group had less tumor weight than the control group(P<0.05).The protein expression of PCNA,pAKT and p STAT3 in the observation group was lower than that in the control group(P<0.05).Compared with the control group,the number of Ki67-positive cells in the observation group decreased,and the number of apoptotic cells increased(P<0.05).Conclusion:Decreasing the levels of pla

关 键 词：宫颈癌 HPV 巨噬细胞集落刺激因子 基质金属蛋白酶9 基质金属蛋白酶组织抑制因子1 增殖 

分 类 号：R-33[医药卫生] R737.33