浅析Cripto—1蛋白和mRNA在食管鳞癌中的表达及其生物学意义论文_刘群

浅析Cripto—1蛋白和mRNA在食管鳞癌中的表达及其生物学意义论文_刘群

刘群

哈尔滨市香坊区人民医院 150036

摘要:目的 探究Cripto-1在食管鳞癌组织中的表达情况及其与食管鳞癌发生、发展的关系。方法 采用免疫组化SP法和原位杂交方法对60例食管鳞癌及60例正常食管黏膜组织进行Cripto-1蛋白和mRNA的检测。结果(1)原位杂交结果显示,Cripto-1 mRNA在食管鳞癌组织中表达的阳性率为63.33%,显著高于正常食管黏膜组织中Cripto-1 mRNA的表达的阳性率(8.33%),两组之间Cripto-1 mRNA表达的差异具有统计学意义(P < 0.05)。(2)免疫组织化学结果表明,Cripto-1蛋白在食管鳞癌组织中表达的阳性率为58.33%,显著高于正常食管黏膜组织中Cripto-1蛋白的表达的阳性率(6.67%),两组之间Cripto-1蛋白表达的差异具有统计学意义(P < 0.05)。(3)对60例食管鳞癌患者的临床病理学参数进行统计分析,结果显示Cripto-1 mRNA和蛋白的表达均与食管鳞癌患者的年龄和性别无关(P > 0.05),但与食管鳞癌的组织学分级、浸润深度、淋巴结转移和TNM分期具有显著的相关性(P < 0.05)。(4)相关性分析结果表明,在60例食管鳞癌标本中,Cripto-1 mRNA阳性表达的38例患者中,Cripto-1蛋白阳性表达患者占31例,而Cripto-1 mRNA阴性表达的22例患者中,Cripto-1蛋白阴性表达占18例。Cripto-1 mRNA和蛋白在食管鳞癌组织中的表达呈正相关关系(γ=0.600,P = 0.000)。结论 食管鳞癌组织中Cripto-1 mRNA和蛋白均呈现高表达,表明Cripto-1高表达可能与食管鳞癌的发生和进展密切相关。

关键词:Cripto-1;食管鳞癌;免疫组化;原位杂交

[Abstract] [Abstract] Objective To investigate the expression of Cripto-1 in esophageal squamous cell carcinoma and its relationship with the occurrence and development of squamous cell carcinoma of the esophagus. Methods immunohistochemical SP and in situ hybridization were used to detect Cripto-1 protein and mRNA in 60 cases of esophageal squamous cell carcinoma and 60 cases of normal esophageal mucosa. Results(1)the results of in situ hybridization showed that the positive expression rate of Cripto-1 mRNA in esophageal squamous cell carcinoma was 63.33%,significantly higher than the positive rate of Cripto-1 expression in normal esophageal mucosa of the mRNA(8.33%),the difference was statistically significant Cripto-1 mRNA expression between the two groups(P < 0.05).(2)the result of immunohistochemistry showed that the positive rate of Cripto-1 protein expression in esophageal squamous cell carcinoma was 58.33%,significantly higher than the positive rate of Cripto-1 protein expression in normal esophageal mucosa(6.67%),the difference of Cripto-1 expression between the two groups had statistical significance(P < 0.05).(3)the clinicopathological parameters of 60 cases of patients with esophageal squamous cell carcinoma were analyzed,the results showed that the expression of Cripto-1 protein and mRNA were not related to the age and sex of patients with esophageal squamous cell carcinoma(P > 0.05),but the histological grade of esophageal squamous cell carcinoma tissue,and the depth of invasion,lymph node metastasis and TNM staging had significant correlation(P < 0.05).(4)correlation analysis showed that in 60 cases of esophageal squamous cell carcinoma,38 patients with positive expression of Cripto-1 mRNA accounted for 31 cases of Cripto-1 protein positive expression,while 38 cases of Cripto-1 mRNA negative expression accounted for 18 cases of Cripto-1 protein negative expression. The expression of Cripto-1 mRNA and protein in the tissues of esophageal squamous cell carcinoma was positively correlated(gamma =0.600,P = 0). Conclusion the high expression of Cripto-1 mRNA and protein in the tissue of esophageal squamous cell carcinoma indicates that the high expression of Cripto-1 may be closely related to the occurrence and progression of esophageal squamous cell carcinoma.

[Key words] Cripto-1;squamous cell carcinoma of the esophagus;immunohistochemistry;in situ hybridization

食管癌分为两种类型,食管腺癌(esophageal adenocarcinoma,EAC)和食管鳞癌(esophageal squamous cell carcinoma,ESCC)。尽管EAC的发生率在一些西方国家持续上升,但是ESCC仍然占主导优势,其主要发生在伊朗的北部地区和北中国地区,大约占总的食管癌病例的90%[1]。Cripto-1,也被称为畸胎癌来源的生长因子-1(teratocarcinoma-derived growth factor-1,TDGF-1),是表皮生长因子(epidermal growth factor,EGF)-cripto-1-FRL-1-cryptic(CFC)蛋白家族的成员[2]。多项研究结果显示,其异常表达可能在肿瘤的发生过程中起到重要作用[3-4]。然而,迄今为止,经大量的文献检索发现Cripto-1与食管鳞癌发生发展的关系国内外尚未见报道,因此在本研究中,笔者采用原位杂交、免疫组织化学联合检测食管鳞癌组织和正常食管黏膜组织中Cripto-1 mRNA和蛋白的表达水平,并分析其与临床病理学参数之间的关系,该研究旨在为进一步研究Cripto-1在食管鳞癌中的功能提供理论依据。

1 材料与方法

1.1 材料

60例食管鳞癌(无坏死癌灶、癌旁3 cm)和60例对应的正常食管黏膜组织标本均取自食管癌发病率较高的地区河南省安阳市肿瘤医院,并经HE染色证实,所有食管鳞癌患者术前均无放疗、化疗和免疫治疗史。将上述所有的标本采用40 g/L的多聚甲醛进行固定,接着进行一系列的常规脱水,石蜡包埋和连续切片(厚度2~4 μm),然后分别用于HE、原位杂交及免疫组织化学染色。兔抗人Cripto-1 mAb购自美国Santa Cruz公司,SP免疫组化试剂盒购自北京中杉金桥生物技术开发公司。原位杂交5'端生物素标记、全硫代修饰探针由北京奥科生物技术有限公司合成,探针序列:TCACAGCCGGGTAGAAATGCCTGAGGAAAGCAGCGGA。

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1.2方法

1.3 结果判定

免疫组化及原位杂交判定方法主要按阳性细胞所占靶细胞总数量百分比及其着色强度进行结果判定[5-6]。(1)按切片中细胞着色深浅评分:0分,细胞无显色;1分,浅黄色;2分,棕黄色;3分,棕褐色。(2)按阳性细胞数占同类细胞数的百分比,<30%为1分,30%~70%为2分,>70%为3分。取(1),(2)两项评分的乘积作为总积分,0~1分为阴性,≥2分为阳性。

1.4 统计学处理

2 结果

2.1 Cripto-1蛋白在食管鳞癌组织中的表达及其与临床生物学行为的关系

利用免疫组织化学技术检测60例食管鳞癌和相对应的正常食管黏膜组织中Cripto-1蛋白的表达。结果表明,Cripto-1 mRNA主要定位于肿瘤细胞的细胞质中,其染色为浅黄色至棕黄色。对Cripto-1蛋白的染色结果进行评估,结果表明,Cripto-1蛋白在食管鳞癌组织中表达的阳性率为58.33%,显著高于正常食管黏膜组织中Cripto-1蛋白的表达的阳性率(6.67%),二组之间Cripto-1蛋白表达的差异具有统计学意义(P < 0.05)。Cripto-1蛋白表达与食管鳞癌的组织学分级、浸润深度、淋巴结转移和TNM分期具有显著的相关性(P < 0.05),提示Cripto-1蛋白可能与食管鳞癌的发生发展过程密切相关。

3 讨论

人的Cripto-1基因最初在人畸胎癌cDNA的文库中鉴定出来,因此也被称为畸胎癌来源的生长因子-1(teratocarcinoma-derived growth factor-1,TDGF-1),其基因定位于染色体3p21.3,是表皮生长因子(epidermal growth factor,EGF)-cripto-1-FRL-1-cryptic(CFC)蛋白家族的成员[2]。最初对于Cripto-1的研究主要集中在其与胚胎的发育、血管生成的调控以及对细胞迁移和浸润能力的影响。然而最近发现其与肿瘤的发生发展密切相关。有研究表明,Cripto-1能从体外诱导正常上皮细胞的转化,促进上皮细胞间质转换,刺激血管生成[7]。最近大量的证据显示,Cripto-1 mRNA和蛋白已经被证实在多种不同类型的肿瘤中高表达,包括乳腺癌、结肠癌、胃癌和卵巢癌等[2,8-9],提示其在肿瘤的发生发展中发挥重要作用。

总之,本研究结果初步表明Cripto-1 mRNA和蛋白均在食管鳞癌组织中呈现高表达,其高表达与食管鳞癌的组织学分级、浸润深度、淋巴结转移和TNM分期均显著相关,进一步研究Cripto-1在食管鳞癌中的功能有望为食管鳞癌的分子靶向治疗提供理论依据。

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论文作者:刘群

论文发表刊物:《健康世界》2018年5期

论文发表时间:2018/5/25

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浅析Cripto—1蛋白和mRNA在食管鳞癌中的表达及其生物学意义论文_刘群
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