An intracellular role for pancreatic bile salt-dependent lipase: evidence for modification of lipid turnover in transfected CHO cells

Josette Le Petit-Thevenin, Nadine Bruneau, Odette Nobili, Dominique Lombardo, Alain Vérine
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引用次数: 9

Abstract

Pancreatic bile salt-dependent lipase (BSDL) hydrolyzes cholesteryl esters, triglycerides and phospholipids. BSDL is also capable of transferring free fatty acid to cholesterol. BSDL has been detected in many cells including fetal and tumor cells, hepatocytes, macrophages and eosinophils and in tissues such as adrenal glands and testes. The enzyme may be secreted or located within subcellular compartments such as the endoplasmic reticulum or the cytosol. Although the role of the secreted enzyme is well documented, that of the intracellular form(s) is still hypothetical. In the present study, we addressed the effects of BSDL on cell lipid metabolism. For that purpose, the cDNA of rat BSDL was transfected into CHO K1 cells (CHO K1-BSDL clone) which were then loaded with [3H]oleic acid. The results demonstrate that the transfected BSDL is secreted; in spite of that, a large fraction of catalytically active BSDL is found in cell lysate. The lipid metabolism of transfected cells is affected and BSDL induces an enhanced incorporation of [3H]oleic acid in cholesteryl esters whereas fatty acid incorporation in phosphatidylcholine is decreased. These effects were particularly important in the cytosol of transfected cells where transfected BSDL preferentially locates. These data suggested that BSDL could be implicated in the cycle of the cellular homeostasis of cholesterol which is particularly affected in tumoral cells leading to cholesteryl ester storage within cytosolic lipid droplets.

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胰腺胆盐依赖性脂肪酶的细胞内作用:在转染CHO细胞中脂质转换的证据
胰胆盐依赖性脂肪酶(BSDL)水解胆固醇酯、甘油三酯和磷脂。BSDL还能将游离脂肪酸转化为胆固醇。BSDL已在胎儿细胞、肿瘤细胞、肝细胞、巨噬细胞、嗜酸性粒细胞等多种细胞以及肾上腺、睾丸等组织中检测到。酶可以分泌或位于亚细胞区室,如内质网或细胞质内。尽管分泌酶的作用已被充分证明,但细胞内形式的作用仍是假设的。在本研究中,我们探讨了BSDL对细胞脂质代谢的影响。为此,将大鼠BSDL的cDNA转染CHO K1细胞(CHO K1-BSDL克隆),然后负载[3H]油酸。结果表明:转染后的BSDL得以分泌;尽管如此,在细胞裂解液中发现了大部分具有催化活性的BSDL。转染细胞的脂质代谢受到影响,BSDL诱导[3H]油酸在胆固醇酯中的掺入增强,而脂肪酸在磷脂酰胆碱中的掺入减少。这些作用在转染细胞的细胞质中尤其重要,转染的BSDL优先定位在细胞质中。这些数据表明,BSDL可能与胆固醇的细胞稳态循环有关,尤其是在肿瘤细胞中受到影响,导致胆固醇酯在细胞质脂滴内储存。
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