Feng Bo, Zheng Min-hua, Ma Jun-jun, Cai Qu, Zhang Yi, Ji Jun, Qu Ying, Li Jian-wen, Lu Ai-guo, Wang Ming-liang, Liu Bing-ya, Zhu Zheng-gang
Department of General Surgery, Ruijin Hospital, Medical College of Shanghai Jiaotong University, Shanghai 200025, China.
Zhonghua Wai Ke Za Zhi. 2006 May 1;44(9):623-7.
To demonstrate expression and single nucleotide polymorphisms (SNP) of human kallikrein 10 (KLK 10) in colorectal cancer (CRC) and to correlate the KLK 10 expression level with clinicopathological factors of CRC.
KLK 10 expression in 63 cases of tumoral and nontumoral colorectal tissues at the mRNA and protein levels were evaluated by quantitative real-time RT-PCR (qRT) and Western blot methods. KLK 10 protein was localized by immunohistochemistry. The KLK 10 genomic DNA from 16 cases of paired normal and cancerous colorectal tissues was PCR-amplified and examined for SNP by direct sequencing.
The KLK 10 mRNA expression was detected by qRT in 61 of 63 (97%) CRC specimens. The KLK 10 expression was much higher in tumor tissue than in the corresponding normal mucosal tissue at the mRNA and protein levels. The KLK 10 mRNA expression level significantly correlated with the lymphatic invasion (P < 0.05) and clinical stage of CRC (P < 0.05). No mutations or polymorphisms were detected in exon 1, 2 and 5 of KLK 10 gene in CRC. A SNP in codon 50 of exon 3, GCC (alanine) to TCC (serine) was identified. The genetic changes of exon 4 were located at codon 106 [GGC (glycine) to GGA (glycine)], codon 112 [ACG (threonine) to ACC (threonine)], codon 141 [CTA (leucine) to CTG (leucine)], and codon 149 [CCG (proline) to CTG (leucine)]. All these SNP were identical in tumor as well as the corresponding normal tissue DNA from the same individuals.
The KLK 10 expression is up-regulated in CRC and higher expression of KLK 10 closely correlate with advanced disease stage, which predicts a poorer prognosis, however, further follow-up study is needed.
研究人组织激肽释放酶10（KLK 10）在结直肠癌（CRC）中的表达及单核苷酸多态性（SNP），并将KLK 10表达水平与CRC的临床病理因素相关联。
采用定量实时逆转录聚合酶链反应（qRT）和蛋白质免疫印迹法，评估63例肿瘤及非肿瘤结直肠组织中KLK 10在mRNA和蛋白质水平的表达。通过免疫组织化学对KLK 10蛋白进行定位。对16例配对的正常和癌性结直肠组织的KLK 10基因组DNA进行PCR扩增，并通过直接测序检测SNP。
qRT检测显示，63例CRC标本中有61例（97%）检测到KLK 10 mRNA表达。在mRNA和蛋白质水平上，肿瘤组织中KLK 10的表达均显著高于相应的正常黏膜组织。KLK 10 mRNA表达水平与CRC的淋巴浸润（P < 0.05）及临床分期（P < 0.05）显著相关。在CRC中，未在KLK 10基因的外显子1、2和5中检测到突变或多态性。在外显子3的第50密码子处发现了一个SNP，即GCC（丙氨酸）突变为TCC（丝氨酸）。外显子4的基因变化位于第106密码子[GGC（甘氨酸）变为GGA（甘氨酸）]、第112密码子[ACG（苏氨酸）变为ACC（苏氨酸）]、第141密码子[CTA（亮氨酸）变为CTG（亮氨酸）]和第149密码子[CCG（脯氨酸）变为CTG（亮氨酸）]。所有这些SNP在同一患者的肿瘤组织及其相应正常组织DNA中均相同。
CRC中KLK 10表达上调，KLK 10的高表达与疾病晚期密切相关，提示预后较差，然而，仍需进一步的随访研究。
