Hathorn R W, Tso C L, Kaboo R, Pang S, Figlin R, Sawyers C, deKernion J B, Belldegrun A
Department of Surgery, UCLA School of Medicine 90024-1738.
Cancer. 1994 Oct 1;74(7):1904-11. doi: 10.1002/1097-0142(19941001)74:7<1904::aid-cncr2820740713>3.0.co;2-b.
Continuous local delivery of interleukin-2 (IL-2) and interferon-alpha (IFN-alpha) via gene transfer appears to be more effective than systemic therapy in preventing the growth of human renal cell carcinoma (RCC) in vitro and in vivo. To understand further if cytokine-gene transfection of RCC could alter certain cellular properties that are associated with the invasive and metastatic potentials of tumor, the authors characterized six cell lines that produce IL-2 and/or IFN-alpha in their expression of intercellular adhesion molecule-1 (ICAM-1) and CD44; binding affinity to extracellular matrix (ECM) components (fibronectin, laminin, type IV collagen, and vitronectin); and preference in forming homotypic aggregation and mRNA levels of c-myc, epidermal growth factor receptor (EGF-R), tumor transforming growth factor-beta (TGF-beta) and type IV collagenase. These six lines were compared with control vector transfected parental R11 line.
The expression of ICAM-1 and CD44 was determined by fluorescence-activated cell sorter (FACS) analysis, the tumor cell binding affinity to ECM components was measured by cell attachment assay, the degree of homotypic aggregation was quantified by cell aggregation assay, and the mRNA levels of c-myc, EGF-R, TGF-beta, and collagenase were analyzed by a quantitative polymerase chain reaction analysis.
Both IL-2-gene- and IFN-alpha-gene-modified R11 exhibited enhanced expression of ICAM-1, suppression of CD44, and decreased binding affinity to ECM components, when compared with the R11-control vector. All cytokine-producing tumor lines showed a decreased preference to form homotypic aggregation. Interferon-alpha gene transfer downregulated c-myc, EGF-R, and type IV collagenase mRNA expression, whereas only the higher producers of IL-2 downregulated TGF-beta mRNA expression. Exogenous IL-2 and/or IFN-alpha treatment of a IFN-alpha-resistant RCC enhanced both HLA class I antigen and ICAM-1 expression and suppressed CD44 expression, but had no effect on tumor growth rate.
The local production of high concentrations of IL-2 and IFN-a at the tumor site may directly alter tumor properties associated with invasive and metastatic phenotypes of RCC. Interleukin-2 and/or IFN-alpha gene therapy may be an effective strategy for treatment of patients with advanced renal cancer.
通过基因转移持续局部递送白细胞介素-2（IL-2）和干扰素-α（IFN-α）在体外和体内预防人肾细胞癌（RCC）生长方面似乎比全身治疗更有效。为了进一步了解RCC的细胞因子基因转染是否会改变与肿瘤侵袭和转移潜能相关的某些细胞特性，作者对六种在细胞间黏附分子-1（ICAM-1）和CD44表达中产生IL-2和/或IFN-α的细胞系进行了表征；对细胞外基质（ECM）成分（纤连蛋白、层粘连蛋白、IV型胶原和玻连蛋白）的结合亲和力；形成同型聚集的偏好以及c-myc、表皮生长因子受体（EGF-R）、肿瘤转化生长因子-β（TGF-β）和IV型胶原酶的mRNA水平。将这六种细胞系与对照载体转染的亲本R11细胞系进行比较。
通过荧光激活细胞分选仪（FACS）分析确定ICAM-1和CD44的表达，通过细胞附着试验测量肿瘤细胞与ECM成分的结合亲和力，通过细胞聚集试验量化同型聚集程度，并通过定量聚合酶链反应分析c-myc、EGF-R、TGF-β和胶原酶的mRNA水平。
与R11对照载体相比，IL-2基因和IFN-α基因修饰的R11均表现出ICAM-1表达增强、CD44表达受抑制以及与ECM成分的结合亲和力降低。所有产生细胞因子的肿瘤细胞系形成同型聚集的偏好均降低。干扰素-α基因转移下调了c-myc、EGF-R和IV型胶原酶mRNA的表达，而只有较高水平产生IL-2的细胞系下调了TGF-β mRNA的表达。用外源性IL-2和/或IFN-α处理对IFN-α耐药的RCC可增强HLA I类抗原和ICAM-1的表达并抑制CD44的表达，但对肿瘤生长速率没有影响。
肿瘤部位高浓度IL-2和IFN-α的局部产生可能直接改变与RCC侵袭和转移表型相关的肿瘤特性。白细胞介素-2和/或干扰素-α基因治疗可能是治疗晚期肾癌患者的有效策略。
