Rivera R T, Pasion S G, Wong D T, Fei Y B, Biswas D K
Laboratory of Molecular Biology, Harvard School of Dental Medicine, Boston, Massachusetts 02115.
J Cell Biol. 1989 Jun;108(6):2423-34. doi: 10.1083/jcb.108.6.2423.
A clonal strain of human lung tumor cells in culture (ChaGo), derived from a bronchogenic carcinoma, synthesizes and secretes large amounts of alpha (alpha) and a comparatively lower level of beta (beta) subunit of the glycoprotein hormone, human chorionic gonadotropin (HCG). ChaGo cells lost their characteristic anchorage-independent growth phenotype in the presence of anti-alpha-HCG antibody. The effect of the antibody was partially reversed by addition of alpha-HCG to the culture medium. ChaGo cells were transfected with an expression vector (pRSV-anti-alpha-HCG), that directs synthesis of RNA complementary to alpha-HCG mRNA. The transfectants produced alpha-HCG antisense RNA which was associated with the reduced level of alpha-HCG. Transfectants also displayed several altered phenotypic properties, including altered morphology, less mitosis, reduced growth rate, loss of anchorage-independent growth, and loss of tumorigenicity in nude mice. Treatment of transfectants with 8,bromo-cAMP resulted in increased accumulation of alpha-HCG mRNA, no change in the level of alpha-HCG antisense RNA, release of the inhibition of [3H]thymidine incorporation, and restoration of anchorage-independent growth phenotype. The overexpression of c-myc, observed in ChaGo cells, was unaffected by the reduced level of alpha-HCG. These results suggest that ectopic synthesis of the alpha subunit of HCG plays a functional role in the transformation of these human lung cells.
一种培养的人肺肿瘤细胞克隆株(ChaGo)源自支气管癌,能合成并分泌大量糖蛋白激素人绒毛膜促性腺激素(HCG)的α亚基,且β亚基的分泌水平相对较低。在抗α-HCG抗体存在的情况下,ChaGo细胞失去了其特征性的不依赖贴壁生长表型。向培养基中添加α-HCG可部分逆转抗体的作用。用一种表达载体(pRSV-抗α-HCG)转染ChaGo细胞,该载体可指导合成与α-HCG mRNA互补的RNA。转染子产生α-HCG反义RNA,这与α-HCG水平降低有关。转染子还表现出几种表型特性的改变,包括形态改变、有丝分裂减少、生长速率降低、失去不依赖贴壁生长以及在裸鼠中失去致瘤性。用8-溴-cAMP处理转染子导致α-HCG mRNA积累增加,α-HCG反义RNA水平无变化,解除对[3H]胸腺嘧啶掺入的抑制,并恢复不依赖贴壁生长表型。在ChaGo细胞中观察到的c-myc过表达不受α-HCG水平降低的影响。这些结果表明,HCGα亚基的异位合成在这些人肺细胞的转化中发挥功能作用。