DSCR1基因与喉咽鳞状细胞癌血管新生关系的研究

发布时间：2018-06-12 15:03

本文选题：DSCR1基因 + 喉咽鳞状细胞癌　；参考：《山东大学》2012年博士论文

【摘要】：目的通过研究喉咽鳞状细胞癌组织与癌旁组织中DSCR1基因蛋白的表达,了解其表达与肿瘤临床特征的关系,探讨DSCR1基因对下咽鳞状细胞癌的生物学行为的影响,为以DSCR1基因为靶点的抗肿瘤治疗提供生物学依据。方法收集94例喉咽鳞状细胞癌病例,有78例同时取癌旁组织,通过免疫组织化学p-V9000法,使用兔抗DSCR1抗体DCT3,检测DSCR1基因蛋白在喉咽鳞状细胞癌与正常癌旁组织中的表达,数据输入SAS8.2软件进行统计学分析,采用卡方检验比较不同性别、年龄、组织学分级、临床分期、部位、生长方式、淋巴结转移以及吸烟史组间表达差异,配对t检验方法对78例同一病人的肿瘤与癌旁组织中基因蛋白的表达差异进行分析,P0.05为有统计学意义。结果肿瘤组织中DSCR1基因蛋白表达阳性率94.9%,癌旁组织中表达阳性率35.9%,有极显著统计学意义(t值23.69,P0.001)；DSCR1基因的表达与组织学分级、TNM分期显著相关(P0.05)；DSCR1基因的表达与与肿瘤的发病年龄、性别、部位、生长方式、淋巴结转移以及吸烟史均无关(P0.05)。结论 1.在喉咽癌肿瘤组织与癌旁组织中,DSCR1基因蛋白的表达存在明显的差异。 2.DSCR1基因与肿瘤的组织学分级有显著关系,分化程度越差,DSCR1基因蛋白的表达阳性率越高,与TNM分期及淋巴结转移的关系有待于进一步研究。 3.在喉咽癌的形成及发展进程中,DSCR1基因必定发挥了重要的作用,可能是此基因的失活或功能失调导致了肿瘤的发展不受控制,并影响了肿瘤的生物学特征,是一个很有前途的肿瘤基因治疗靶点。目的通过检测喉咽癌组织与癌旁组织中微血管密度数值,研究血管因素与肿瘤侵袭性间的相关性。并与前期测定的DSCR1基因表达之间做相关性分析,讨论DSCR1基因与喉咽癌血管生成之间的关系,探讨DSCR1基因对喉咽癌的发生、血管新生等生物学行为特性的影响。方法使用免疫组化PV9000方法,在与第一部分相同的94例喉咽鳞状细胞癌病例,通过CD34抗体染色标记血管内皮细胞,计算微血管密度(MVD)值,通过与临床资料进行统计学分析,探讨MVD与各种临床病理参数之间的关系,并通过前期测定的肿瘤组织中DCT3抗体标记的DSCR1基因表达与MVD之间做相关性分析,MVD结果以均数±标准差表示,两组间比较采用t检验(方差齐),三组以上样本间关系采用单因素方差分析,P0.05表示差异有显著性。MVD与DSCR1表达相关性检验用Spearman相关分析。结果在喉咽癌组织中MVD平均为17.57±10.61条/400倍视野,癌周正常组织为12.61±7.67条/400倍视野,经统计学分析差异显著(P0.05)。MVD与肿瘤颈淋巴结转移的关系研究发现,转移组中MVD平均为18.96±11.63条/400倍视野,而非转移组为16.17±9.61条/400倍视野,经统计学分析差异显著(P0.05)；MVD与喉咽癌的病理分级的关系发现,高分化组的MVD平均为16.29±9.98条/400倍视野,中分化组为18.58±10.35条/400倍视野,低分化组为20.48±13.61条/400倍视野,三组比较差异显著(P0.05)。对瘤内血管密度MVD与肿瘤临床分期的相关性研究表明,TNM分期Ⅰ～Ⅱ组MVD平均为16.13±9.61/400倍视野,而Ⅲ～Ⅳ组为18.43±11.61条/400倍视野,经统计学分析差异显著(P0.05)；本实验表明MVD与患者的发病年龄、性别、肿瘤生长部位、生长方式及吸烟史等均无明显相关性(P0.05)；在喉咽癌肿瘤组织中随着MVD的升高,DSCR1基因的表达也逐渐增强,结果具有显著的统计学意义(P0.0001),且两者之间为正相关的关系(r=0.698)。结论 1.喉咽癌组织中MVD与肿瘤的组织学分级、TNM分期、淋巴结转移有关。说明肿瘤不同的生物学特征伴随不同的微血管密度表达,可以通过检测MVD判断喉咽癌的预后,以及高MVD的喉咽癌患者行预防性颈淋巴结清扫及术后的放、化疗等综合治疗是必要的。 2.在喉咽癌鳞状细胞癌,随着DSCR1基因的表达活性增加,肿瘤微血管密度的也增强,两者存在正相关的关系,证实了DSCR1基因与肿瘤血管生成的密切相关性,说明DSCR1基因可以影响喉咽癌的侵袭、转移等过程,是很有前途的肿瘤治疗靶点。目的了解VEGF-C mRNA及DSCRl mRNA在喉咽鳞癌组织及癌旁组织中的表达水平,进一步验证VEGF及DSCRl基因与肿瘤的关系,并将VEGF-C mRNA表达水平与DSCR1mRNA水平做相关性分析,探讨DSCR1基因与喉咽癌血管生成之间的关系。方法通过RT-PCR方法检测DSCR1mRNA及VEGF-C mRNA在43例喉咽鳞癌组织及21例癌旁组织中的表达情况,在不同组织学分级、临床分期、淋巴结转移之间进行比较,并对二者的表达进行相关性分析。结果43例喉咽癌中35例可检测到DSCRl mRNA阳性,占81.39%,21例正常癌旁组织中8例可检测DSCR1mRNA阳性,占38.10%,喉咽鳞状细胞癌DSCR1mRNA表达的平均相对光密度值为0.91±0.10,明显高于正常癌旁组织0.61±0.08,差异具有统计学意义(P0.05)；DSCR1mRNA的表达在不同的组织学分级、及淋巴结转移均有显著差异(P0.05)。43例喉咽癌32例可检测到VEGF-C mRNA,占74.42%,21例正常组织中6例可检测至VEGF-C mRNA,占28.57%,癌周组织平均相对灰度值为(0.56±0.19),癌内组织平均相对灰度值为(1.21±0.41),二组之间比较差异均有统计学意义(P0.05)。43例肿瘤组织中DSCR1mRNA表达与VEGF-C mRNA表达的相关性,经Spearman相关性分析,两者之间成正相关。结论 1.在喉咽癌肿瘤组织与癌旁组织中,DSCR1mRNA的表达存在明显的差异。DSCR1与不同的肿瘤组织学分级及淋巴结转移有显著关系,此基因与肿瘤的生物学行为密切相关。 2. VEGF-C mRNA的表达从癌周组织到癌内组织相对灰度值呈增高趋势,有显著差异,说明VEGF-C与癌症的发生发展有密切关系。 3.在喉咽癌肿瘤组织中DSCR1mRNA表达与VEGF-CmRNA表达呈正相关,DSCR1基因与喉咽鳞状细胞癌的血管生成存在密切关系。
[Abstract]:Objective to study the expression of DSCR1 gene protein in the tissues of larynx squamous cell carcinoma and paracancerous tissue, to understand the relationship between the expression of the tumor and the clinical features of the tumor, and to explore the effect of the DSCR1 gene on the biological behavior of the hypopharyngeal squamous cell carcinoma, and to provide a biological basis for the anti tumor treatment of the target based on the DSCR1 base.
Methods 94 cases of hypopharynx squamous cell carcinoma were collected and 78 cases were collected simultaneously. The expression of DSCR1 gene protein in hypopharynx squamous cell carcinoma and normal para cancerous tissue was detected by immunohistochemistry p-V9000 method and Rabbit anti DSCR1 antibody DCT3. The data were analyzed by SAS8.2 software, and the same sex was compared with the chi square test. No, age, histological grade, clinical stage, location, growth mode, lymph node metastasis and smoking history, the difference in the expression of gene protein in 78 cases of the same patient's tumor and para cancerous tissue was analyzed by paired t test, and P0.05 was statistically significant.
Results the positive rate of DSCR1 gene protein expression in tumor tissues was 94.9%, the positive rate of the Para Para tissue was 35.9%, with significant statistical significance (t value 23.69, P0.001); the expression of DSCR1 gene was significantly correlated with the histological grade, TNM staging (P0.05); the expression of the DSCR1 gene and the age of the tumor, sex, location, growth mode, lymph node, and lymph node Metastasis and the history of smoking were not related (P0.05).
conclusion
1. there was a significant difference in the expression of DSCR1 protein in tumor tissues and adjacent tissues of hypopharyngeal carcinoma.
There is a significant relationship between the 2.DSCR1 gene and the histological grade of the tumor. The worse the degree of differentiation, the higher the expression of DSCR1 gene protein, the relationship with TNM staging and lymph node metastasis need to be further studied.
3. in the process of the formation and development of hypopharynx cancer, the DSCR1 gene is bound to play an important role. It may be that the inactivation or dysfunction of the gene may lead to the uncontrolled development of the tumor, and affect the biological characteristics of the tumor. It is a promising target for the treatment of tumor genes.
Objective to investigate the correlation between vascular factors and tumor invasiveness by detecting the microvascular density in the tissues of hypopharyngeal carcinoma and adjacent tissues, and to analyze the correlation between the DSCR1 gene expression and the relationship between the DSCR1 gene and the angiogenesis of the hypopharynx cancer, and explore the occurrence of the DSCR1 gene and the angiogenesis of the hypopharynx. The effects of biological behavior characteristics.
Methods using immunohistochemical PV9000 method, 94 cases of larynx squamous cell carcinoma the same as the first part were used to mark vascular endothelial cells by CD34 antibody staining, and the value of microvascular density (MVD) was calculated. The relationship between MVD and various clinicopathological parameters was analyzed by statistical analysis with clinical data. The correlation analysis between the DSCR1 gene expression of DCT3 antibody markers in the tissue and MVD was analyzed, and the results of MVD were expressed with mean standard deviation. The two groups were compared with t test (Fang Chaqi), and the relationship between the three groups was analyzed by single factor analysis of variance. P0.05 indicated that the difference was significant in the correlation test of.MVD and DSCR1 expression by Spearman correlation analysis.
Results the average MVD of MVD in the hypopharynx cancer tissue was 17.57 + 10.61 /400 times of vision, and the normal tissue of the carcinoma was 12.61 + 7.67 times of /400 fold. The statistical difference was significant (P0.05) and the relationship between the tumor neck lymph node metastasis and the MVD was 18.96 + 11.63 /400 times of vision in the transfer group, while the non metastasis group was 16.17 + 9.61 /400 times. The visual field was statistically significant (P0.05). The relationship between the MVD and the histopathological classification of hypopharynx found that the average MVD of the high differentiation group was 16.29 + 9.98 /400 times of vision, the middle differentiation group was 18.58 + 10.35 /400 times of vision, the low differentiation group was 20.48 + 13.61 /400 times of vision, and the three groups were significantly different (P0.05). The intra tumor vascular density MVD and swelling were found. The correlation study of the clinical staging of the tumor showed that the average MVD of TNM stage I to II group was 16.13 + 9.61/400 fold, while the group III to IV was 18.43 + 11.61 /400 times of /400, and the difference was significant (P0.05). The experiment showed that there was no significant correlation between MVD and the age of the patients, the sex, the tumor growth position, the growth mode and the smoking history. Sex (P0.05); the expression of DSCR1 gene was gradually increased with the increase of MVD in the tumor tissues of hypopharynx cancer, and the results had significant statistical significance (P0.0001), and the relationship between them was positively related (r=0.698).
conclusion
The histological grading, TNM staging and lymph node metastasis in 1. hypopharynx cancer tissues are related to the different biological characteristics of the tumor with different microvascular density expressions. The prognosis of the laryngynx cancer can be judged by the detection of MVD, as well as the comprehensive treatment of prophylactic neck lymph node dissection and postoperative radiotherapy and chemotherapy for high MVD hypopharynx cancer patients. It is necessary.
2. in the squamous cell carcinoma of hypopharynx cancer, with the increase of the expression of DSCR1 gene, the tumor microvessel density is also enhanced. There is a positive correlation between the two. It confirms the close correlation between the DSCR1 gene and the angiogenesis of tumor. It shows that the DSCR1 gene can affect the invasion and metastasis of the larynx cancer, and it is a promising target for cancer treatment.
Objective to investigate the expression of VEGF-C mRNA and DSCRl mRNA in hypopharynx squamous cell carcinoma tissue and para cancer tissue, to further verify the relationship between VEGF and DSCRl genes and tumor, and to analyze the correlation between the expression level of VEGF-C mRNA and the level of DSCR1mRNA, and to explore the relationship between the DSCR1 gene and the angiogenesis of the hypopharynx cancer.
Methods the expression of DSCR1mRNA and VEGF-C mRNA in 43 cases of hypopharynx squamous cell carcinoma and 21 para cancerous tissues were detected by RT-PCR method. The comparison of the histological grade, clinical stage and lymph node metastasis was carried out, and the expression of the two were analyzed.
Results in 43 cases of larynx cancer, 35 cases were detected to be DSCRl mRNA positive, accounting for 81.39%. 8 cases of normal para cancerous tissues in 21 cases were detected DSCR1mRNA positive, accounting for 38.10%. The average relative light density of DSCR1mRNA expression in hypopharynx squamous cell carcinoma was 0.91 + 0.10, which was significantly higher than that of normal para cancerous tissue (0.61 + 0.08), and the difference was statistically significant (P0.05); DSCR1mRNA There were significant differences in histological grade and lymph node metastasis (P0.05) in 32 cases of hypopharynx cancer in.43, VEGF-C mRNA was detected in 74.42%. In 21 normal tissues, 6 cases were detected to VEGF-C mRNA, 28.57%, and the average relative gray value of the tissue was (0.56 + 0.19), and the average relative gray value in the tissues was (1.21 + 0.41), and two groups were between the two groups. The difference was statistically significant (P0.05) the correlation between the expression of DSCR1mRNA and the expression of VEGF-C mRNA in the tumor tissues of.43, and the correlation between them was positively correlated with the Spearman correlation analysis.
conclusion
1. there is a significant difference in the expression of DSCR1mRNA in the tumor tissues and para cancerous tissues of the hypopharynx cancer..DSCR1 is closely related to the histological grade and lymph node metastasis of different tumors. This gene is closely related to the biological behavior of the tumor.
The expression of 2. VEGF-C mRNA increased from the peritacid tissue to the relative gray level in the tissues, which showed significant differences, indicating that VEGF-C was closely related to the development of cancer.
3. there was a positive correlation between the expression of DSCR1mRNA and the expression of VEGF-CmRNA in hypopharyngeal carcinoma tissues. DSCR1 gene is closely related to angiogenesis in hypopharyngeal squamous cell carcinoma.
【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R739.65

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