背景与目的:前列腺癌抗原3(prostate cancer antigen 3,PCA3)作为一种长链非编码RNA(lncRNA)已被证实在前列腺癌中高度特异性表达,暗示其可能在前列腺癌中发挥重要作用,为深入研究,我们将完整的PCA3基因转入真核表达载体,并构建慢病毒包装系统。方法:从前列腺癌细胞株LNCaP细胞中提取总RNA,运用重叠延伸方法扩增到PCA3基因,测序正确后定向接入真核表达载体pCDH-CMV-MCS-EF1-copGFP,酶切鉴定并测序正确后进行慢病毒包装和滴度测定。结果:经PCR鉴定及测序,PCA3表达质粒序列与Gene Bank序列进行Blast比对分析,同源性为99.8%,PCA3慢病毒滴度测定为2×108。结论:PCA3成功插入真核表达载体并完成了慢病毒包装,为深入研究PCA3基因在前列腺细胞中的作用奠定了基础,进而为探索前列腺癌的治疗提供了新的途径。
Prostate cancer gene 3 （PCA3, also known as DD3） is a new biomarker that could improve the accuracy of prostate cancer diagnosis. It is a great biomarker with fairly high specificity and sensitivity. The incidence of prostate cancer is rising steadily in most countries. The commonly used prostate-specific antigen （PSA） test once gave people hope for early diagnosis of prostate cancer. However, the low specificity of the PSA test has resulted in a large number of unnecessary biopsies and overtreatment. During the past decade, many new prostate cancer biomarkers have been found. Among these, PCA3 is the most promising. Due to its great performance in distinguishing prostate cancer from other prostate conditions, PCA3 could likely be applied for early diagnosis of prostate cancer, patient follow-up, prognosis prediction, and targeted therapy. After years of research, we have obtained some knowledge about the sequence of PCA3 gene. We have also determined the relationship between PCA3 and the proliferation of prostate cancer cells and learned some information about how PCA3 affects tumor-related genes and proteins. A PCA3 score has been created, and it has been used in a variety of studies. Some researchers have even applied PCA3 to targeted therapy and obtained a good effect in vitro. This review describes the current state of research, and explores the future prospects for PCA3.
