摘要
目的对正常乳腺组织和癌组织KLK6 mRNA进行定量,分析其表达与临床病理特征的关系,并探讨乳腺癌细胞株MCF-7中KLK6基因被沉默后细胞增殖及凋亡的变化。方法应用实时荧光定量PCR检测48例乳腺癌切除组织中KLK6基因mRNA的表达量,并分析KLK6 mRNA的表达量与ER、PR及肿瘤转移情况的相关性。以体外化学合成的siRNA干扰乳腺癌细胞株MCF-7中KLK6基因的表达,分析细胞周期、细胞凋亡及细胞形态的变化。结果KLK6 mRNA在73%(35/48)的乳腺癌组织中表达水平增高,明显高于正常组织(P<0.01),有淋巴结转移的患者癌组织中的表达水平明显低于未转移组(P<0.05);ER(+)组低于ER(-)组(P<0.05);PR(+)组与PR(-)组差异不明显(P>0.05);与乳腺癌相关基因CerbB-2的表达无相关性(P>0.05)。MCF-7细胞中KLK6基因表达沉默后,S期细胞增加13.23%,细胞凋亡无明显变化。结论KLK6 mRNA在乳腺癌组织表达增高,并与ER状态及肿瘤转移情况有关。体外MCF-7细胞株和体内移植瘤KLK6基因表达沉默后肿瘤细胞生长加快。提示KLK6基因表达受雌激素下调的影响,是促乳腺癌细胞增殖的抑制因子,从而为乳腺癌的防治提供了新思路。
objective To quantify the expression of kallikrein gene 6 （KLK6） in malignant and benign breast tissues and to statistically analyze the relationship between KLK6 expression levels and the clinico-pathological variables in patients with breast cancer. Method Paired breast tissue samples of cancerous and corresponding non-cancerous tissues were obtained from 48 patients with breast cancer who underwent surgical resection. Quantitative analysis of KLK6 mRNA expression was done using real-time quantitative reverse transcription-PCR （RT-PCR）. The relationship between KLK6 expression intensity and various clinico-pathological variables was analyzed. In addition, the technique of small interfering RNA-mediated gene silencing was used to suppress the KLK6 expression in MCF-7, and to investigate the effects of KLK6 on cell proliferation rate and cell cycle. Results KLK6 mRNA over expression was found in cancerous tissues in 35 out of 48 patients （73%） as compared with normal breast tissue. The mean expression level of KLK6 mRNA in cancerous tissues was significantly higher than that in non-cancerous tissues （P〈0. 01）. Elevated KLK6 expression was lower in patients with metastasis and significantly negatively correlated with metastasis （P〈0.05） and ER status （P〈0. 05）. Furthermore, it was found that when KLK6 gene was silenced with KLK6 small interfering RNA effectively increased the cell proliferation rate and cell population in the S phase. Conclusions The present study indicates that KLK6 mRNA expression is significantly higher in cancerous than in non-cancerous breast tissues, and the high expression of KLK6 mRNA is correlated with metastasis and ER status. It is also suggested that KLK6 may possibly be a suppressor gene for breast cancer.
出处
《解放军医学杂志》
CAS
CSCD
北大核心
2007年第10期1085-1087,共3页
Medical Journal of Chinese People's Liberation Army
基金
全军医学科研"十一五"计划面上项目(06MA101)
