循环肿瘤细胞分型及FAM172A检测在结直肠癌中的临床价值研究

发布时间：2018-05-10 08:34

本文选题：结直肠癌 + 循环肿瘤细胞　；参考：《南方医科大学》2017年硕士论文

【摘要】：目的:本文利用多重RAN原位杂交方法,对结直肠癌患者外周血循环肿瘤细胞进行分型及FAM172A检测,分析其表达与患者肿瘤侵润深度、淋巴结转移、远处转移、TNM分期等临床特征参数之间的关系,研究不同类型循环肿瘤细胞的转移潜能,探讨循环肿瘤细胞分型与FAM172A在结直肠癌诊疗中的临床价值。方法:1.采集2015年6月至2015年12月南方医科大学珠江医院普通外科45例结直肠癌患者外周血5ml,采集同期收治10例结直肠良性肿物患者外周血5ml。将红细胞裂解过滤,通过直径8 μm的纳米膜系统富集循环肿瘤细胞,然后利用4组特异性核酸探针进行多重RAN原位杂交,通过不同颜色荧光信号来鉴定循环肿瘤细胞的分型及FAM172A表达情况。2.统计学分析:本实验数据采用Spss20.0软件进行统计学分析,其中CTCs、FAM172A与患者临床特征参数的关系采用x2检验,计量资料的组间比较采用非参数检验,当P0.05表示为差异有统计学意义。结果:1.运用纳米膜技术与多重RAN原位分析技术的检测方法,对循环肿瘤细胞分为eCTCs(上皮型)、bCTCs(混合型)、mCTCs(间质型);45例结直肠癌患者检测CTCs阳性率为62.2%,mCTCs检测阳性率为57.8%,而对照组患者未检测CTCs存在,两组CTCs阳性率比较差异有统计学意义(P0.05)。2.循环肿瘤细胞阳性率与结直肠癌大多数临床特征无明显相关关系(P0.05),但间质型循环肿瘤细胞与结直肠癌发生脉管侵润及远处转移呈显著相关(P0.05)。3.FAM172A在CTCs总体检出率为60.7%,其中上皮型CTCs为56.3%,混合型CTCs 为 58.6%,而间质型 CTCs 为 68.8%。4.在循环肿瘤细胞阳性的结直肠癌患者中,FAM172A表达率与肿瘤侵润深度及Ki-67水平呈显著相关(P0.05),在间质型循环肿瘤细胞阳性的结直肠癌患者中,FAM172A表达率与肿瘤侵润深度及脉管侵润呈显著相关(P0.05)。5.在Ⅱ期结直肠癌患者中,高危组患者中mCTCs阳性率及FAM172A表达阳性率均明显高于低危组患者的mCTCs阳性率及FAM172A表达阳性率,但两组比较差异无统计学意义(P0.05)。需增加样本量检测对此结果加以验证。结论:1.基于纳米膜技术与多重RAN原位分析技术的检测方法,可以有效对结直肠癌患者外周血循环肿瘤细胞进行分型,包括上皮型、混合型及间质型,并且在不同类型的循环肿瘤细胞中FAM172A表达率不同,其中在间质型循环肿瘤细胞最高。2.间质型循环肿瘤细胞可能与结直肠癌血行转移相关,可能参与肿瘤进展过程。3.FAM172A可能参与结直肠癌的侵袭与转移过程,与结直肠癌进展相关。4.联合检测循环肿瘤细胞分型及FAM172A,有望成为预测结直肠癌患者预后的潜在有效生物学指标。
[Abstract]:Objective: to study the expression of circulating tumor cells in peripheral blood of patients with colorectal cancer and to analyze their expression with tumor invasion depth and lymph node metastasis by in situ hybridization with multiple RAN. To study the metastatic potential of different circulating tumor cells and to explore the clinical value of circulating tumor cell classification and FAM172A in the diagnosis and treatment of colorectal cancer. Method 1: 1. From June 2015 to December 2015, the peripheral blood of 45 patients with colorectal cancer in Zhujiang Hospital of Southern Medical University and 5 ml of peripheral blood of 10 patients with benign colorectal tumor were collected. The circulating tumor cells were enriched with RBC cleavage and filtration through a nanofilm system with diameter of 8 渭 m, and then multiplex RAN in situ hybridization was performed using four groups of specific nucleic acid probes. The cell types and FAM172A expression of circulating tumor cells were identified by different color fluorescence signals. Statistical analysis: the data of this experiment were analyzed by Spss20.0 software. The relationship between CTCsfam 172A and the clinical characteristic parameters of patients was tested by x2 test. The non-parametric test was used in the comparison of measurement data. The difference was statistically significant when P0.05 was expressed. The result is 1: 1. The detection method of nanofilm and multiplex RAN in situ analysis was used. The positive rate of CTCs was 62.2% in 45 patients with colorectal cancer, while no CTCs was detected in the control group. There was a significant difference between the two groups in the positive rate of CTCs. There was no significant correlation between the positive rate of circulating tumor cells and most clinical features of colorectal cancer. However, there was a significant correlation between circulating tumor cells of interstitial type and vascular invasion and distant metastasis of colorectal cancer. 3. FAM172A was detected in 60.7 patients with CTCs. The epithelial type of CTCs was 56.3%, the mixed type of CTCs was 58.6%, and the interstitial type of CTCs was 68.8%. 4. The expression rate of FAM172A was significantly correlated with the depth of tumor invasion and the level of Ki-67 in colorectal cancer patients with positive circulating tumor cells. The expression rate of FAM172A was significantly correlated with the depth of tumor invasion and the depth of tumor invasion in patients with colorectal cancer with positive interstitial circulating tumor cells. There was a significant correlation between vascular invasion and embellishment (P0.05. 5). The positive rate of mCTCs and FAM172A expression in high risk group was significantly higher than that in low risk group, but there was no significant difference between the two groups. This result needs to be verified by increasing sample size. Conclusion 1. Based on the technique of nanomembrane and multiplex RAN in situ analysis, it can be used to type circulating tumor cells in peripheral blood of colorectal cancer patients, including epithelial type, mixed type and interstitial type. The expression rate of FAM172A was different in different types of circulating tumor cells, and the highest expression rate of FAM172A was found in stromal circulating tumor cells. Interstitial circulating tumor cells may be associated with blood metastasis of colorectal cancer. 3. FAM172A may be involved in the invasion and metastasis of colorectal cancer, and may be related to the progression of colorectal cancer. Combined detection of circulating tumor cell types and FAM172A is expected to be a potential biological marker for predicting the prognosis of colorectal cancer patients.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R735.34

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