College of Veterinary Medicine, Inner Mongolia Agricultural University, Hohhot, China; Key Laboratory of Basic Veterinary Medicine of Inner Mongolia Autonomous Region, Hohhot, China.
Eur Rev Med Pharmacol Sci. 2021 Feb;25(3):1410-1424. doi: 10.26355/eurrev_202102_24849.
The incidence of thyroid cancer is rising globally. Most patients progress slowly, but some patients develop lymph node and distant metastasis earlier, and their prognosis is poor. Therefore, early diagnosis and warning of malignancy are very meaningful for such patients. SAS1B gene is a newly discovered protein expressed on the surface of mature egg cells and has metalloendopeptidase activity. We aimed at exploring whether SAS1B is involved in the occurrence of thyroid cancer, and at providing evidence for early diagnosis and targeted therapy of thyroid cancer.
In this study, a rabbit anti-human SAS1B polyclonal antibody was prepared by gene recombination technology. The indirect ELISA method was used to detect the SAS1B protein expression in the serum of 69 patients with thyroid cancer and 55 normal controls, and the relevant pathological factors were analyzed. Immunohistochemistry and PCR technology were used to investigate the expression levels of SAS1B protein and mRNA in 30 thyroid cancer tissues and 23 control thyroid tissues.
The titer of SAS1B recombinant antibody was 1:51200. The expression of SAS1B in the serum of patients with thyroid cancer was higher than that in the normal control group (p<0.01). The antibody had a good sensitivity in serum detection of cancer patients (p=0.008<0.01), the linear regression analysis result was that the expression of SAS1B gene was related to tumor envelope invasion and lymph node metastasis (p=0.003<0.01, p=0.003<0.01), and it was irrelevant to the patient's gender, age, tumor mass size, number of cancer foci, pathological stage, etc. (p>0.05). The results of immunohistochemistry showed that SAS1B protein was mainly located in the cytoplasm and membrane of thyroid cancer cells. The expression intensity in thyroid cancer tissues was higher than that in control tissues (p<0.05), but it was not expressed in normal thyroid tissues. Antibodies showed a good sensitivity that was used to detect thyroid cancer tissues (p=0.000<0.01). The results of ordinary PCR detection using thyroid cancer tissue and control thyroid tissue showed that the amplification products of the three domains (N-terminal, C-terminal and catalytic domain) of the SAS1B gene showed high expression in thyroid cancer tissue. q-PCR results showed that the expression of SAS1B gene in thyroid cancer and control thyroid tissue was higher than that in control group (p<0.05), and the genes of Aurora A and BARD1 related to centrosome replication and DNA replication forks protection during the proliferation were highly expressed in thyroid cancer tissue. The study results suggested that SAS1B was involved in the carcinogenesis of thyroid cancer. The Hum_mPLoc.2.0 software, PSORT Ⅱ software and UniProt software were used to predict that SAS1B protein had secretory protein properties.
The above data indicate that the SAS1B gene is closely related to the process of thyroid cancer and can serve as a good tumor marker that can be used for early diagnosis and early warning of thyroid malignancy.
甲状腺癌的发病率在全球范围内呈上升趋势。大多数患者进展缓慢，但有些患者较早出现淋巴结和远处转移，预后较差。因此，早期诊断和恶性预警对这类患者具有重要意义。SAS1B 基因是一种新发现的成熟卵母细胞表面表达的蛋白，具有金属内肽酶活性。我们旨在探讨 SAS1B 是否参与甲状腺癌的发生，并为甲状腺癌的早期诊断和靶向治疗提供依据。
本研究采用基因重组技术制备兔抗人 SAS1B 多克隆抗体。采用间接 ELISA 法检测 69 例甲状腺癌患者和 55 例正常对照者血清中 SAS1B 蛋白的表达情况，并分析相关病理因素。采用免疫组化和 PCR 技术检测 30 例甲状腺癌组织和 23 例对照甲状腺组织中 SAS1B 蛋白和 mRNA 的表达水平。
SAS1B 重组抗体的效价为 1:51200。甲状腺癌患者血清中 SAS1B 的表达高于正常对照组（p<0.01）。该抗体对癌症患者的血清检测具有良好的灵敏度（p=0.008<0.01），线性回归分析结果表明，SAS1B 基因的表达与肿瘤包膜浸润和淋巴结转移有关（p=0.003<0.01，p=0.003<0.01），与患者的性别、年龄、肿瘤大小、癌症灶数量、病理分期等无关（p>0.05）。免疫组化结果显示，SAS1B 蛋白主要位于甲状腺癌细胞的细胞质和膜中。甲状腺癌组织中表达强度高于对照组（p<0.05），但在正常甲状腺组织中不表达。抗体对甲状腺癌组织的检测具有良好的灵敏度（p=0.000<0.01）。用甲状腺癌组织和对照甲状腺组织进行普通 PCR 检测的结果显示，SAS1B 基因的三个结构域（N 端、C 端和催化结构域）的扩增产物在甲状腺癌组织中高表达。q-PCR 结果显示，甲状腺癌组织和对照甲状腺组织中 SAS1B 基因的表达均高于对照组（p<0.05），与中心体复制和 DNA 复制叉保护相关的 Aurora A 和 BARD1 基因在甲状腺癌组织中高表达。研究结果表明，SAS1B 参与了甲状腺癌的发生。Hum_mPLoc.2.0 软件、PSORT Ⅱ软件和 UniProt 软件预测 SAS1B 蛋白具有分泌蛋白特性。
上述数据表明，SAS1B 基因与甲状腺癌的发生过程密切相关，可作为甲状腺恶性肿瘤早期诊断和早期预警的良好肿瘤标志物。
