Genetic integrity of transforming growth factor beta (TGF-beta) receptors in cervical carcinoma cell lines: loss of growth sensitivity but conserved transcriptional response to TGF-beta

Int J Cancer. 1998 Aug 12;77(4):620-5. doi: 10.1002/(sici)1097-0215(19980812)77:4<620::aid-ijc23>3.0.co;2-8.

Abstract

Transforming growth factor beta (TGF-beta) exerts an inhibitory effect on the growth of most epithelial cell types, and the loss of responsiveness to this growth inhibition has been implicated in the development of a variety of human cancers. The genetic alteration of TGF-beta receptors is known to play a critical role in this escape from growth regulation. We asked whether there is a correlation between TGF-beta sensitivity and the genetic status of TGF-beta type I and type II receptors (RI and RII, respectively) in human cervical carcinoma cell lines. Among 8 cell lines examined, 3 (ME-180, C-33A and HeLaS3) showed resistance to TGF-beta and 3 (SiHa, CaSki and HeLa229) showed minimal response to the growth inhibitory effect of TGF-beta; the other cell lines (HeLa and HT-3) were sensitive. Northern blot analysis revealed that the RII mRNA was not expressed in 2 TGF-beta-resistant cell lines (ME-180 and C-33A) but was expressed in the other cell lines. Southern blot analysis of RI and RII revealed a homozygous deletion of the entire TGF-beta RII gene in the cell line ME-180. We then asked whether the other TGF-beta-resistant or refractory cell lines had microsatellite instability and/or poly-adenine tract mutations of RII. We also checked for point mutations in the individual exons of the entire RII using polymerase chain reaction-single-strand conformational polymorphism (PCR-SSCP). Although C-33A exhibited poly-adenine microsatellite instability, its RII gene showed no signs of mutation. The molecular integrity of the TGF-beta, receptors in all cell lines, except ME-180 and C-33A, could be confirmed by examining the distinct transcriptional induction of plasminogen activator inhibitor-1 (PAI-1), p21(WAF1/CIP1) and, in some cases, the accompanying downregulation of c-myc in response to TGF-beta. Our observations, taken together, indicate that inactivation of the RII contributes to the resistance to TGF-beta of some cervical carcinoma cell lines. Loss of or attenuated sensitivity to TGF-beta growth inhibition in other cells may be attributed to the disruption of distal components in the TGF-beta signal pathway, but not to the receptor system.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Southern
  • Cell Division / drug effects
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins / metabolism
  • DNA, Neoplasm / analysis
  • Female
  • HeLa Cells / drug effects
  • Humans
  • Microsatellite Repeats / genetics
  • Plasminogen Activator Inhibitor 1 / metabolism
  • RNA, Messenger / metabolism
  • Receptors, Transforming Growth Factor beta / genetics*
  • Transcriptional Activation
  • Transforming Growth Factor beta / pharmacology
  • Trinucleotide Repeat Expansion / genetics
  • Tumor Cells, Cultured / drug effects
  • Uterine Cervical Neoplasms / genetics*
  • Uterine Cervical Neoplasms / metabolism
  • Uterine Cervical Neoplasms / pathology

Substances

  • CDKN1A protein, human
  • Cyclin-Dependent Kinase Inhibitor p21
  • Cyclins
  • DNA, Neoplasm
  • Plasminogen Activator Inhibitor 1
  • RNA, Messenger
  • Receptors, Transforming Growth Factor beta
  • Transforming Growth Factor beta