Guan Xiaoxiang, Li Mingfang, Fan Leming, Chen Qi
Atherosclerosis Research Center, Nanjing Medical University, Nanjing, Jiangsu, 210029 P. R. China.
Zhonghua Yi Xue Yi Chuan Xue Za Zhi. 2003 Apr;20(2):138-42.
To investigate low density lipoprotein receptor (LDLR) function and gene mutation in Chinese patients with familial hypercholesterolemia(FH).
Lymphocytes were isolated from 10 ml anticoagulated peripheral blood of the patients, then a flow-cytometric method (FCM) with 1,1'-dioctadecyl-3,3,3', 3-tetramethylindocarbocyanine perchlorate labelled low density lipoproetin (DiI-LDL) was used to identify the function of LDLR on the surface of lymphocytes. Genomic DNA was isolated from whole blood of FH patients and analyzed by PCR-single strand conformation polymorphism (SSCP) and nucleotide sequencing methods.
Defects of binding and uptaking of LDLR were identified by FCM in 2 FH patients in one family, and their parents were examined in the present study. Then they were analyzed genetically. The detected mutation was a deletion of A, which caused a frame shift in codon 297 of exon 6 and introduced a beforehand stop codon in codon 369.
A novel mutation of LDL receptor gene was detected by the combination of FCM and PCR-SSCP methods.
研究中国家族性高胆固醇血症（FH）患者的低密度脂蛋白受体（LDLR）功能及基因突变情况。
从患者10ml抗凝外周血中分离淋巴细胞，然后采用1,1'-二辛基-3,3,3',3-四甲基吲哚羰花青高氯酸盐标记的低密度脂蛋白（DiI-LDL）流式细胞术（FCM）鉴定淋巴细胞表面LDLR的功能。从FH患者全血中提取基因组DNA，采用聚合酶链反应-单链构象多态性（PCR-SSCP）和核苷酸测序方法进行分析。
通过FCM在一个家族的2例FH患者中鉴定出LDLR结合和摄取缺陷，并对其父母进行了本研究检查，随后进行了基因分析。检测到的突变是A的缺失，导致外显子6第297密码子移码，并在第369密码子引入提前终止密码子。
通过FCM和PCR-SSCP方法联合检测到一种新的LDL受体基因突变。
