扩散加权成像及自噬相关蛋白表达在宫颈癌中的临床价值研究

发布时间：2018-04-24 05:24

本文选题：宫颈癌 + 细胞自噬　；参考：《郑州大学》2015年博士论文

【摘要】：研究背景宫颈癌发病率及死亡率居全球女性恶性肿瘤的第二位,在我国则居于女性生殖系统恶性肿瘤的第一位,因此,如何采取有效措施降低宫颈癌的发病率与死亡率,已经成为重要的社会公共健康问题。细胞自噬是实现细胞的代谢和细胞器的新旧交替,维持细胞自身稳定的生物学过程。自噬及相关蛋白的表达情况,与多种疾病特别是恶性肿瘤的发生、发展密切相关。Beclin1是自噬体在形成过程中的一个重要蛋白,它可以促进自噬前体的形成,调控自噬体的形成和成熟过程;p62是一种选择性自噬受体,主要参与细胞内膜膜流、DNA修复、介导泛素化蛋白等自噬降解过程,是反映自噬活性的标记性蛋白;LC-3由LC-3-Ⅰ和LC-3-II组成,主要参与自噬体的形成。由于LC-3-II的含量与自噬小泡的数量成正比,因此LC-3(主要是LC-3-II)常作为评价自噬活性的重要指标。进行自噬相关蛋白表达如Beclin1、p62及LC-3蛋白表达与宫颈癌的相关性研究,可以进一步揭示宫颈癌发生和发展机制,为宫颈癌的预后判断、有效预防及寻找宫颈癌治疗新靶点提供依据。扩散加权成像(DWI)是目前唯一能够在体外测定活体组织内水分子扩散情况的影像学检查技术,通过对宫颈癌DWI图像表观扩散系数(ADC)值的定量分析,可以较为准确地反映有关宫颈癌肿瘤的功能性改变,但能否用于对宫颈癌预后的预测和评价方面,尚缺乏相关的研究性证据。本研究采用免疫组化方法探讨自噬相关蛋白Beclin1、p62和LC-3在正常子宫颈组织、宫颈上皮内瘤变(CIN)、子宫颈鳞状细胞癌组织中的阳性表达情况,分析上述三种蛋白与宫颈癌临床病理因素之间的关系,以及对宫颈癌生存和预后的影响。同时利用磁共振扩散加权成像(DWI)对不同分化程度子宫颈鳞状细胞癌的ADC值进行测定,探讨自噬相关蛋白与功能磁共振定量参数间的关系,为宫颈癌的转化医学研究提供新的思路和方法。目的观察beclin1、p62及lc-3在正常子宫颈、cin和宫颈癌中的阳性表达情况,分析上述三种自噬相关蛋白的表达与宫颈癌临床病理因素之间的关系,以及对宫颈癌生存和预后的影响。测定高、中、低分化子宫颈鳞状细胞癌患者的adc值并进行分析比较,同时研究宫颈癌自噬相关蛋白表达与adc值间的相关性。由此探讨扩散加权成像及自噬相关蛋白表达在宫颈癌中的价值。材料与方法采用免疫组化sp法对宫颈癌组(子宫颈鳞状细胞癌)标本191例、cin组标本63例和正常子宫颈组标本32例进行beclin1、p62及lc-3蛋白阳性表达的标记和半定量评分比较,分析上述三种蛋白表达与患者临床病理因素之间的关系,以及对宫颈癌组患者总生存时间和无进展生存时间的影响。同时将宫颈癌组患者dwi影像资料后处理所得adc值与自噬相关蛋白表达进行分析比较。结果1.beclin1蛋白定位于细胞膜及细胞质中,在正常子宫颈组、cin组及宫颈癌组的阳性表达率分别为31.3%、36.5%和19.4%,具有统计学差异(x2=19.323,p=0.000)。beclin1蛋白在宫颈癌组阳性表达的评分值低于正常子宫颈组及cin组(p0.01,p0.001),而且阳性表达与患者的年龄、肿瘤分化、figo分期、sccag值、肿瘤直径、肌层浸润深度、手术切缘、死亡无统计学差异(p值均0.05),与淋巴结转移和复发有统计学差异(p值均0.05)。2.p62蛋白定位于细胞质和细胞核中,在正常子宫颈组、cin组及宫颈癌组的阳性表达率分别为65.6%、65.1%、73.8%,具有统计学差异(x2=9.617,p0.01)。p62蛋白在宫颈癌组阳性表达的评分值高于正常子宫颈组与cin组(p值均0.01),并且阳性表达与患者的年龄、肿瘤分化、figo分期、肿瘤直径、肌层浸润深度、手术切缘、死亡无统计学差异(p值均0.05),与sccag值、淋巴结转移和复发有统计学差异(p值均0.05)。3.lc-3蛋白定位于细胞质和细胞核中,在正常子宫颈组、cin组及宫颈癌组的阳性表达率分别为84.4%、73.0%、50.3%,具有统计学差异(x2=36.291,p=0.000)。lc-3蛋白在宫颈癌组阳性表达的评分值低于正常子宫颈组及cin组(P0.001,P0.01),而且与患者的年龄、FIGO分期、SCCAg值、肿瘤直径、手术切缘、死亡无统计学差异(P值均0.05),与肿瘤分化、淋巴结转移、肌层浸润深度及复发有统计学差异(P值均0.05)。4.肿瘤分化、淋巴结转移、LC-3表达是影响总生存时间的危险因素(P值均0.05),年龄、FIGO分期、SCCAg值、肿瘤直径、肌层浸润深度、手术切缘、Beclin1表达和p62表达与总生存时间无统计学差异(P值均0.05);SCCAg值、淋巴结转移、Beclin1表达、p62表达和LC-3表达是影响宫颈癌组患者无进展生存时间的危险因素(P值均0.05);年龄、肿瘤分化、FIGO分期、肿瘤直径、肌层浸润深度和手术切缘与无进展生存时间无统计学差异(P值均0.05)。5.宫颈癌组患者中高分化宫颈癌患者的平均ADC值中分化宫颈癌患者的平均ADC值低分化宫颈癌患者的平均ADC值(P0.05)。6.在宫颈癌组中,高分化患者的LC-3阳性表达与ADC值无统计学差异,中、低分化患者中LC-3阳性表达与ADC值间有统计学差异(P0.05)。结论1.Beclin1蛋白在子宫颈鳞状细胞癌组织中呈现低表达,与肿瘤的淋巴结转移和复发具有统计学差异,且是影响无进展生存时间的危险因素,说明Beclin1蛋白可能与宫颈癌的发生及发展密切相关。2.p62蛋白在子宫颈鳞状细胞癌组织中呈现高表达,与肿瘤的淋巴结转移、SCCAg值和肿瘤的复发具有统计学差异,且是影响无进展生存时间的危险因素,说明p62蛋白可能与宫颈癌的发展及预后相关。3.LC-3蛋白在子宫颈鳞状细胞癌组织中呈现低表达,与肿瘤的分化、淋巴结转移、肌层浸润深度和复发具有统计学差异,且是影响总生存时间及无进展生存时间的危险因素,说明LC-3蛋白与宫颈癌的发生、发展及预后密切相关。4.ADC值测定不仅可用于判断子宫颈鳞状细胞癌的分化程度,而且与LC结合可能可以用于宫颈癌的预后评估。
[Abstract]:Background the incidence and mortality of cervical cancer are the second most malignant tumors in the world. In our country, it is the first of the female reproductive system malignant tumors. Therefore, how to take effective measures to reduce the incidence and mortality of cervical cancer has become an important social public health problem. Cell autophagy is the realization of cell metabolism and the development of cell autophagy. The new and old alternation of organelles, the biological process of maintaining cell self stability. The expression of autophagy and related proteins, and the development of.Beclin1 is an important protein in the formation of autophagic bodies, which can promote the formation of autophagic precursors and regulate the formation and maturation of autophagic bodies. Process; p62 is a selective autophagy receptor, which is mainly involved in the cell membrane flow, DNA repair, and ubiquitination protein, which is a marker protein reflecting autophagic activity; LC-3 is composed of LC-3- I and LC-3-II and is mainly involved in the formation of autophagic. As the content of LC-3-II is proportional to the number of autophagic vesicles, LC-3 (principal) If LC-3-II) is often used as an important indicator of autophagic activity, the correlation between the expression of autophagy related protein, such as Beclin1, p62 and LC-3 protein expression and cervical cancer, can further reveal the mechanism of cervical cancer development and development, and provide a basis for the prognosis of cervical cancer, the effective prevention and search for new targets for cervical cancer treatment. Right imaging (DWI) is the only imaging technique that can determine the diffusion of water molecules in living tissues in vitro. By quantitative analysis of the apparent diffusion coefficient (ADC) value of the DWI image of cervical cancer, the functional alteration of cervical cancer can be accurately reflected, but it can be used to predict and evaluate the prognosis of cervical cancer. In this study, there is still a lack of relevant research evidence. In this study, the immunohistochemical method was used to investigate the positive expression of autophagy related protein Beclin1, p62 and LC-3 in normal cervical tissue, cervical intraepithelial neoplasia (CIN), cervical squamous cell carcinoma, and analysis of the relationship between the three proteins and the clinicopathological factors of cervical cancer, and the relationship between these proteins and the clinicopathological factors of cervical cancer, and The effects on the survival and prognosis of cervical cancer. At the same time, the ADC value of cervical squamous cell carcinoma with different degree of differentiation was measured by magnetic resonance diffusion weighted imaging (DWI). The relationship between autophagy related proteins and quantitative parameters of functional magnetic resonance (fMRI) was explored to provide new ideas and methods for the research of the transformation medicine of cervical cancer. The purpose of this study was to observe Beclin1, p62 And the positive expression of LC-3 in normal cervix, CIN and cervical cancer. The relationship between the expression of the three autophagy related proteins and the clinicopathological factors of cervical cancer, and the influence on the survival and prognosis of cervical cancer. The ADC value of the patients with high, medium and low differentiated cervical squamous cell carcinoma was analyzed and compared, and the study of the uterus was also studied. The correlation between the expression of autophagy related protein and the value of ADC in cervical cancer. The value of DWI and autophagy related protein expression in cervical cancer was discussed. Materials and methods were treated by immunohistochemical SP method in 191 specimens of cervical cancer group (cervical squamous cell carcinoma), 63 cases in group CIN and 32 cases of normal cervix group for Beclin1, p62 and l C-3 protein positive expression and semi quantitative comparison were used to analyze the relationship between the three protein expressions and the patients' clinicopathological factors, as well as the total survival time and the progression free survival time of the patients with cervical cancer group. At the same time, the ADC value of the DWI image data in the cervical cancer group was expressed with the autophagy related protein. Results 1.beclin1 protein was located in the cell membrane and cytoplasm. The positive expression rate in the normal cervical group, the CIN group and the cervical cancer group were 31.3%, 36.5% and 19.4% respectively. The statistical difference (x2=19.323, p=0.000) was lower than the normal cervix group and the CIN group (P0.01, p0.00) in the cervical cancer group. 1) and positive expression with the patient's age, tumor differentiation, FIGO staging, SCCAg value, tumor diameter, myometrium infiltration depth, surgical margin, death without statistical difference (P value is 0.05), and lymph node metastasis and recurrence (both P 0.05).2.p62 protein located in the cytoplasm and nucleus, in the normal cervix group, CIN group and cervix. The positive expression rates of the cancer group were 65.6%, 65.1% and 73.8% respectively. The positive expression of.P62 protein in the cervical cancer group was higher than that of the normal cervical group and the CIN group (P value of 0.01), and the positive expression was with the age of the patients, the tumor differentiation, the FIGO stage, the tumor diameter, the depth of the myometrium infiltration, the surgical margin and death. Statistical difference (P value was 0.05), and SCCAg value, lymph node metastasis and recurrence were statistically different (P value was 0.05).3.lc-3 protein located in cytoplasm and nucleus, in normal cervical group, CIN group and cervical cancer group positive expression rate was 84.4%, 73%, 50.3%, with statistical difference (x2=36.291, p=0.000).Lc-3 protein in cervical cancer group The score of positive expression was lower than that of normal cervix group and CIN group (P0.001, P0.01), and there was no statistical difference between the patients' age, FIGO staging, SCCAg value, tumor diameter, surgical margin and death (P value was 0.05). There were statistical differences with tumor differentiation, lymph node metastasis, myometrium infiltration depth and recurrence (P value was 0.05).4. tumor differentiation, lymph node turn. LC-3 expression was a dangerous factor affecting the total survival time (P value was 0.05). Age, FIGO staging, SCCAg value, tumor diameter, myometrium infiltration depth, surgical margin, Beclin1 expression, p62 expression and total survival time were not statistically different (P value was 0.05); SCCAg, lymph node metastasis, Beclin1 expression, p62 expression and LC-3 expression were affected by cervical cancer. The risk factors of non progressive survival time (P value was 0.05); age, tumor differentiation, FIGO staging, tumor diameter, myometrium infiltration depth, surgical margin and non progressive survival time (P value of all 0.05). The average ADC value of highly differentiated cervical cancer patients in the.5. cervical cancer patients was lower than the average ADC value of differentiation of cervical cancer patients. The average ADC value (P0.05).6. of cervical cancer patients was in the cervical cancer group. There was no statistical difference between the positive expression of LC-3 and the value of ADC in the highly differentiated patients. In the patients with low differentiation, there was a statistical difference between the positive expression of LC-3 and the ADC value (P0.05). Conclusion the expression of 1.Beclin1 protein in the squamous cell carcinoma of the cervix is low, with the lymph node metastasis of the tumor and the metastasis of the tumor. The recurrence has statistical difference, and it is a risk factor that affects the time of no progress. It is suggested that Beclin1 protein may be closely related to the occurrence and development of cervical cancer and the high expression of.2.p62 protein in cervical squamous cell carcinoma tissue, and the lymph node metastasis of tumor, SCCAg value and tumor recurrence have statistical difference, and it has no effect on the recurrence of tumor. The risk factors for the development of the survival time indicate that the p62 protein may be associated with the development and prognosis of cervical cancer and the low expression of.3.LC-3 protein in the squamous cell carcinoma of the cervix, which is statistically different from the tumor differentiation, lymph node metastasis, the depth and recurrence of the myometrium, and is a danger to the total survival time and the progression free survival time. The risk factor indicates that LC-3 protein is closely related to the occurrence, development and prognosis of cervical cancer, and the determination of.4.ADC value can not only be used to judge the degree of differentiation of cervical squamous cell carcinoma, but also with LC may be used to evaluate the prognosis of cervical cancer.

【学位授予单位】：郑州大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R737.33;R445.2

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