p53 gene alternation in squamous cell carcinoma of the esophagus detected by PCR-cold SSCP analysis.

L S Wang, K C Chow, C C Liu, J H Chiu
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Abstract

Mutations of the p53 tumor suppressor gene play an important role in the development of common human malignancies. Previous reports revealed that the frequencies of p53 alternations in esophageal carcinoma varied from 26% to 87%. The clinical significance of p53 alternations is still disputed. In the present study, we used polymerase chain reaction--"cold" single-strand conformation polymorphism (PCR-"cold" SSCP)--to investigate p53 genetic alternations in 63 surgical specimens of esophageal squamous cell carcinoma (ESC). Our experiments showed that the optimum buffer temperature for "cold" SSCP analysis was 14 degrees C while the PCR products were around 200-300 bp in size. Among 63 tumorous samples, p53 alternations was detected in 47 tumors, or an incidence rate of 74.6%. For nontumorous mucosal samples, the incidence of p53 alternations was 55.5% (35/63 samples). Additionally, p53 alternations occurred most frequently at exon 6 (50.8%), followed by exon 7 (33.3%), exon 8 (17.5%) and exon 5 (12.7%). Multiple genetic alternations (> or = 2 exons) between p53 exons 5-8 in the same examined samples were found in 21 (33.3%) of 63 tumors, and in 8 (12.7%) of 63 nontumorous mucosal specimens. Our results further showed that p53 alternations did not correlate with age, depth of tumor invasion, lymph node metastasis, tumor stage, cell differentiation or lymphovascular invasion in ESC (P > 0.05). Moreover, the survival rate in patients with p53 alternations was similar to that in patients without p53 alternations (P > 0.05). In conclusion, PCR-"cold" SSCP is a rapid and sensitive method for identifying p53 genetic alternations. p53 genetic alternations occur with a relatively high incidence for ESC, but p53 abnormality has no impact on prognosis.

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pcr -冷SSCP法检测食管鳞状细胞癌中p53基因的改变。
p53肿瘤抑制基因的突变在人类常见恶性肿瘤的发生发展中起着重要作用。先前的报道显示,食管癌中p53突变的频率从26%到87%不等。p53突变的临床意义仍有争议。在本研究中,我们采用聚合酶链反应-“冷”单链构象多态性(PCR-“冷”SSCP)-研究了63例食管鳞状细胞癌(ESC)手术标本中p53基因的改变。我们的实验表明,“冷”SSCP分析的最佳缓冲温度为14℃,而PCR产物的大小约为200-300 bp。在63例肿瘤样本中,有47例肿瘤检测到p53改变,发生率为74.6%。在非肿瘤粘膜样本中,p53改变的发生率为55.5%(35/63个样本)。此外,p53突变最常发生在外显子6(50.8%),其次是外显子7(33.3%),外显子8(17.5%)和外显子5(12.7%)。63例肿瘤中的21例(33.3%)和63例非肿瘤粘膜标本中的8例(12.7%)发现了p53外显子5-8之间的多重遗传改变(>或= 2外显子)。结果显示,ESC患者p53表达变化与年龄、肿瘤浸润深度、淋巴结转移、肿瘤分期、细胞分化及淋巴血管浸润无关(P > 0.05)。p53突变组与非p53突变组生存率比较,差异无统计学意义(P > 0.05)。总之,PCR-“冷”SSCP是一种快速、灵敏的检测p53基因突变的方法。p53基因改变在ESC中发生率较高,但p53异常对预后无影响。
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