精子相关抗原9（SPAG9）在宫颈癌及宫颈上皮内瘤变中的表达及意义

发布时间：2018-01-13 23:16

  本文关键词：精子相关抗原9（SPAG9）在宫颈癌及宫颈上皮内瘤变中的表达及意义　出处：《山东大学》2014年硕士论文　论文类型：学位论文

  更多相关文章： 精子相关抗原9(SPAG9) 宫颈癌 宫颈上皮内瘤变 诊断

【摘要】：背景： 宫颈癌是女性第三常见的恶性肿瘤,在恶性肿瘤死亡率中占第四位,严重威胁女性健康。并且,全球超过85%宫颈癌新发及死亡病例发生于发展中国家。宫颈癌的发生与发展是一个多阶段、多步骤、复杂的过程,从癌前病变发展为宫颈癌可经历十余年时间,故宫颈癌是一种可预防、甚至可治愈的疾病。尽管,近些年来国内外逐渐普及宫颈癌的筛查工作,虽已使宫颈癌的发病率、死亡率有明显下降,但是因宫颈液基细胞学检查特异性以及敏感性不高,并且阴道镜和宫颈活检等属创伤性检查,也存在漏诊的可能。另外,宫颈病变的术前及预后监测并不完善,目前尚没有一种可靠的肿瘤标志物检测。因此,寻找一种能为宫颈病变的早期筛查、临床诊断及术后检测提供帮助的新型肿瘤标志物显得尤为重要。目的： 检测精子相关抗原9(SPAG9)在宫颈癌、宫颈上皮内瘤变(cervical intraepithelial neoplasia, CIN)组织中的表达,并检测SPAG9在宫颈病变患者血清内的免疫原性,评估其作为新宫颈癌血清标志物的潜在价值。方法： 1、研究对象：子宫颈病变组织及正常宫颈组织选取自山东大学附属省立医院2009年至2012年宫颈手术或活检患者标本。新鲜子宫颈癌标本40例；宫颈上皮内瘤变(CIN)患者手术后石蜡标本35例、新鲜标本7例；对照组选取20例正常宫颈组织(因非宫颈病变而手术)。另外,40例子宫颈癌、40例CIN患者血样均取自须行手术患者,另收集20例子宫颈癌术后返院化疗患者、20例子宫肌瘤患者及10例健康女性血样做为对照。 2、研究方法： (1)应用免疫组织化学(IFC)方法,明确SPAG9在各宫颈组织中的表达定位,通过对平均光密度值的检测,比较正常宫颈组织、宫颈上皮内瘤变组织以及子宫颈癌组织中SPAG9的表达差异。 (2)应用逆转录-聚合酶链反应(RT-PCR)方法检测正常宫颈组织、CIN组织及宫颈癌组织中SPAG9的表达,并通过半定量分析各组间差异。 (3)通过Western Blot方法检测正常宫颈组织、CIN组织及宫颈癌组织中SPAG9的蛋白表达,进行半定量分析。 (4)应用酶联免疫吸附试验(ELISA)检测10例健康女性、20例子宫肌瘤患者、40例宫颈上皮内瘤变患者、40例宫颈癌患者以及20例宫颈癌术后返院化疗、无复发患者的血清标本中SPAG9浓度差异。 结果： 1、免疫组织化学结果显示,SPAG9主要定位于肿瘤细胞的细胞质和细胞膜上,间质中则极少量表达或不表达。SPAG9在正常宫颈组、宫颈上皮内瘤变组及宫颈癌组中的表达量依次增强,且各组间表达的差异均有统计学意义。但SPAG9的表达与宫颈癌临床病理学特征,无论在病理类型、组织学分型、临床分期或有无淋巴结转移等方面的表达差异均无统计学意义。 2、RT-PCR结果显示SPAG9在正常宫颈组织中极少表达或无表达,而在宫颈癌组织中表达量明显增高。通过对SPAG9mRNA在不同宫颈组织中表达的半定量分析显示,在宫颈癌组织中平均表达量为0.815±0.553,而在正常宫颈组织中为0.008±0.001,两组之间差异具有统计学意义。 3、Western Blot检测SPAG9蛋白在宫颈癌及正常宫颈组织中的表达趋势,与免疫组织化学以及RT-PCR的结果一致。半定量分析SPAG9蛋白在宫颈癌组织中平均表达量为0.929±0.908,正常宫颈组织中平均表达量仅为0.044±0.032,差异有统计学意义。 4、ELISA结果示10名健康女性血清中SPAG9平均血清浓度3.2343±1.0579ng/mL,20例子宫肌瘤患者中为3.3561±0.7426ng/mL,两组间差异无统计学意义。而在40例宫颈上皮内瘤变及40例宫颈癌患者血清中SPAG9的平均表达量分别为6.7015±3.3101ng/mL、15.1404±10.3839ng/mL,宫颈癌组高于CIN组高于子宫肌瘤及健康女性组,且各组间的差异均有统计学意义。另外,在20例宫颈癌术后返院化疗、且无复发的患者血清中SPAG9平均浓度为3.6107±1.5069,较术前有所下降。结论： 1、在宫颈癌和CIN组织中SPAG9的蛋白和mRNA均有高表达,且宫颈癌组高于CIN组高于正常宫颈组,三组间差异均有统计学意义。 2、宫颈癌及CIN患者血清中SPAG9表达量均高于正常健康女性和子宫肌瘤患者,且宫颈癌术后、未复发患者血清浓度较术前有所下降。 3、SPAG9作为一个新的生物学标志物,可为宫颈癌及宫颈上皮内瘤变的早期诊断、术后检测、随诊治疗提供指导意义。
[Abstract]:Background:
Cervical cancer is the third most common female malignant tumors, accounting for fourth in the mortality of malignant tumor, a serious threat to women's health. And, more than 85% of cervical cancer and deaths occur in developing countries. The occurrence and development of cervical cancer is a multi-stage, multi-step and complex process, from the development of precancerous lesions for cervical cancer can be more than ten years of experience in the Imperial Palace, cervical cancer is a preventable and treatable disease. Even though, in recent years at home and abroad gradually spread screening of cervical cancer, although the incidence of cervical cancer, mortality rate decreased significantly, but due to cervical cytology specificity and sensitivity is not high, and colposcopy and cervical biopsy is a traumatic examination, there is the possibility of missed diagnosis. In addition, cervical lesions before surgery and prognosis monitoring is not perfect, there is not a reliable tumor marker detection. Therefore, it is very important to find a new type of tumor marker for early screening of cervical lesions, clinical diagnosis and postoperative detection.
Detection of sperm associated antigen 9 (SPAG9) in cervical cancer, cervical intraepithelial neoplasia (cervical intraepithelial, neoplasia, CIN) expression, and detection of SPAG9 in serum of patients with cervical lesions in immunogenicity, evaluate its potential value as a new cervical cancer serum markers:
1, the object of study: cervical lesions and normal cervical tissue were selected from the affiliated Provincial Hospital of Shandong University from 2009 to 2012. Cervical biopsy or surgical specimens from patients with fresh cervical cancer specimens in 40 cases; cervical intraepithelial neoplasia (CIN) in 35 cases of paraffin embedded specimens of patients after surgery, 7 cases of fresh specimens from 20 cases of normal control group; the cervical tissue (for non cervical lesions and surgery). In addition, 40 cases of cervical carcinoma, 40 cases of CIN patients blood samples were taken from the patients with surgery should be performed, another collection of chemotherapy of 20 cervical cancer patients after hospital, 20 cases of uterine leiomyoma and 10 cases of healthy women were as control.
2, research methods:
(1) immunohistochemical localization (SPAG9) was applied to identify the expression and location of IFC in cervical tissues. The expression of SPAG9 in normal cervical tissues, cervical intraepithelial neoplasia tissues and cervical cancer tissues was compared by measuring the mean optical density.
(2) reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of SPAG9 in normal cervical tissues, CIN tissues and cervical cancer tissues.
(3) the Western Blot method was used to detect the protein expression of SPAG9 in the normal cervical tissues, CIN tissues and cervical cancer tissues, and the semi quantitative analysis was performed.
(4) by enzyme linked immunosorbent assay (ELISA) was detected in 10 cases of healthy women, 20 patients with uterine myoma, 40 cases with cervical intraepithelial neoplasia, 40 cases of cervical cancer patients and 20 cases of cervical cancer hospital after chemotherapy, the SPAG9 concentration difference of serum samples in patients with no recurrence.
Result锛，

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