Characterization of estrogen-responsive epithelial cell lines and their infectivity by genital Chlamydia trachomatis.
Chlamydial attachment and infectivity in vitro and ascending disease and sequelae in vivo have been reported to be enhanced/modulated by estrogen. Endometrial carcinoma cell lines Ishikawa and HEC-1B and the breast cancer lines MCF-7 and HCC-1806 were examined for Chlamydia trachomatis E infectivity. Estrogen receptor (ER) presence was confirmed by Western blot and qRT-PCR analyses. FACS analysis was used to determine the percent of plasma membrane-localized ERs (mERs), and their activity was tested by estrogen binding and competitive estrogen antagonists assays. Chlamydiae grew in all cell lines with HEC (90%) >> MCF-7 (57%)>Ishikawa (51%) >> HCC-1806 (20%). The cell line ER isoform composition was re-defined as: ERalpha + ERbeta + for MCF-7, HCC-1806 and Ishikawa; and ERbeta only for HEC-1B. HeLa cells were also tested and found to express ERbeta, but not ERalpha. A small percentage of both ERs were surface-exposed and functionally active. The endometrium-predominant ERbeta isoform was found in all cell lines, including those most representative of the common sites of C. trachomatis infection. Thus, the role of chlamydial attachment/infectivity will now be analyzed in ERbeta+and-isogenic HEC-1B cells.
Guseva
Natalia V
NV
Department of Microbiology, J.H. Quillen College of Medicine, East Tennessee State University, Box 70579, VA#1-Rm. 141, Johnson City, 37614, USA. guseva@etsu.edu
Dessus-Babus
Sophie C
SC
Whittimore
Judy D
JD
Moore
Cheryl G
CG
Wyrick
Priscilla B
PB
D015153:Blotting, Western; D002460:Cell Line; D002462:Cell Membrane / Q000737:chemistry; D002692:Chlamydia trachomatis / Q000254:growth & development*; D003593:Cytoplasm / Q000382:microbiology / Q000648:ultrastructure; D004847:Epithelial Cells / Q000382:microbiology* / Q000648:ultrastructure; D004967:Estrogens / Q000032:analysis / Q000502:physiology*; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D002479:Inclusion Bodies / Q000382:microbiology / Q000648:ultrastructure; D018613:Microscopy, Confocal; D012333:RNA, Messenger / Q000032:analysis; D011960:Receptors, Estrogen / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D004967:Estrogens; D012333:RNA, Messenger; D011960:Receptors, Estrogen