TRANSFORMATI ON OF HUMAN KI DNEY PROXI MAL TUBULE CELLS BY r as - CONTAI NI NG RETROVI RUSES I mpl i cat i ons f or Tumor Pr ogr essi on BY D . M . NANUS, S. A. D. EBRAHI M, N . H . BANDER, * F. X. REAL, L . M . PFEFFER, T J . R. SHAPI RO, AND A. P ALBI NO Fr om t he Memori al Sl oan- Ket t er i ng Cancer Cent er , * Cor nel l Uni ver si t y Medi cal Cent er , and I The Rockef el l er Uni ver si t y, New Yor k, New Yor k 10021 Numer ous st udi es have i mpl i cat ed vari ous r ol es in car ci nogenesi s f or dom i nant - act i ng oncogenes, act i vat ed by mut at i on, chr omosomal r ear r angement , i nser t i on of a near by promot er el ement , or gene ampl i f i cat i on [ 1- 5] . Ot her st udi es have i ndi - cat ed t hat t umori geni ci t y behaves as a r ecessi ve t r ai t , in t hat dom i nant el ement s ( suppr essor genes) mus t first be i nact i vat ed (e.g ., by gene or chr omosomal del et i on, r ear r angement or mut at i on) in or der f or a cel l to become neopl ast i c ( 6- 8) . Pr es um- abl y, nor mal pr ogeni t or cel l s woul d r equi r e a speci f i c ser i es of gene act i vat i ons and/ or i nact i vat i ons to compl et e t he t r ansf or mat i on pr ocess . Mor eover , dependi ng upon t he tumor t ype and t he di f f er ent i at i on pr ogr amof t he cel l , it is l i kel y t hat t hi s ser i es of changes woul d di f f er . The det er m i nat i on of a cont r i but i ng r ol e f or oncogenes in t he sequence of st eps r equi r ed f or mal i gnant t r ansf or mat i on woul d r equi r e, t her e- f or e, an abi l i t y t o di r ect l y t est t he ef f ect s of oncogenes on normal di f f er ent i at ed di pl oi d pr ogeni t or cel l s of a par t i cul ar t umor t ype ( 9) . Human r enal cel l s pr ovi de a sui t abl e syst em f or exam i ni ng t he t emporal sequence of oncogene- r el at ed ef f ect s f or sever al r easons . Fi r st , cel l s f r om bot h r enal cancer s and nor mal human ki dney can be cul - t ur ed, t her eby enabl i ng a di r ect compar i son of t he nor mal and mal i gnant pheno- t ypes ( 10- 13) . Second, i mmunof l uorescent st ai ni ng with mAbs gener at ed agai nst gl y- copr ot ei ns, gl ycol i pi ds, and bl ood gr oup ant i gens of nor mal and mal i gnant r enal cel l s demonst rat e t hat - 90%of cul t ur ed nor mal human ki dney cel l s and vi r t ual l y al l r enal cel l car ci nomas ( as di st i ngui shed f r om t r ansi t i onal cel l car ci nomas of t he ki dney) ar e of proxi mal t ubul e der i vat i on ( 10, 14- 20) . Thus, t r ansf or mat i on exper i - ments with nor mal proxi mal t ubul e ( PT) ' cel l s woul d presumabl y di r ect l y r el at e to event s occur r i ng in vi vo. Thi r d, speci f i c quant i t at i ve and qual i t at i ve changes in proxi mal t ubul ar ant i gens t hat occur upon neopl ast i c t r ansf or mat i on of t he PT cel l Thi s i nvest i gat i on was suppor t ed by U . S. Publ i c Heal t h Ser vi ce gr ant CA- 37907, awar ded by t he Na- t i onal Cancer I nst i t ut e, Depar t ment of Heal t h and Human Ser vi ces ; gr ant GM- 36716; and by t he Char l es H. Revson Foundat i on . D . M . Nanus is a Cl i ni cal Schol ar in Bi omedi cal Resear ch suppor t ed by t he Char l es H. Revson Foundat i on, a r eci pi ent of an Amer i can Phi l osophi cal Soci et y Dal and Fel l owshi p, and an Amer i can Soci et y of Cl i ni cal Oncol ogy 1988 Young I nvest i gat or s Awar d. L . M . Pf ef f er is a Leukem ia Soci et y of Amer i ca Schol ar . 1 Abbr evi at i ons used i n t hi s paper : CALLA, c ommon acut e l ymphobl ast i c l eukem ia ; CB, compl et e buf f er ; CM, condi t i oned ser um- f r ee medi um ; DAB, di am i nobenzi di ne ; EGF, epi dermal gr owt h f act or ; MuLV, muri ne l eukem ia vi r us ; NRK, nor mal r at ki dney ; PT, proxi mal t ubul e ; TGF, t r ansf or m i ng gr owt h f act or ; TLC, t hi n- l ayer chr omat ogr aphy. J . Ex p. Men. © The Rockef el l er Uni ver si t y Pr ess zyxwvutsrqponmlkjihgfedcbaZYXWVUTSRQPONMLKJIHGFEDCBA " 0022- 1007/ 89/ 03/ 953/ 20 $2 . 00 Vol ume 169 Mar ch 1989 953- 972 953 on July 24, 2015 jem.rupress.org Downloaded from Published March 1, 1989