Retinoic acid receptors in retinoid responsive ovarian cancer cell lines detected by polymerase chain reaction following reverse transcription

Abstract

Summary The growth inhibitory effects of all-trans and 13-cis retinoic acid (RA) and of the synthetic retinoids TTNPB, TTNPB-ethylester and TTNN were studied on seven human epithelial ovarian cancer cell lines and one ovarian teratocarcinoma cell line. Six of seven ovarian adenocarcinoma cell lines were inhibited in their growth by RA and by synthetic retinoids in a dose dependent manner. No response to these substances was observed for the ovarian teratocarcinoma cell line. The knowledge that RA and retinoids exert their action on the cells via nuclear receptors led us to examine the expression of RAR-a, -,B and -mRNA by these cell lines by polymerase chain reaction following reverse transcription. All cell lines expressed RAR-x and -mRNA and six of the eight cell lines were found to express additionally RAR-P mRNA, among them the ovarian teratocarcinoma cell line. Our data indicate that there was no direct association between the presence of RAR subtype transcripts and the response to retinoids in ovarian cancer cell lines.Retinoic acid (RA) is a morphogenic compound involved in vertebrate development (Eichele, 1989) and it plays a major role in epithelial cell growth and cellular differentiation (Sporn et al., 1984). RA induces differentiation of mouse embryonal carcinoma cells in vitro and influences the development of the regenerating amphibian limb (for review see de Luca, 1991).It has also been shown that retinoids cause growth inhibition in many hyperproliferating cell lines, a feature that makes the compounds of fundamental interest as antitumour agents. Retinoids prevent the development of cancer of the skin and are effective as agents in the treatment of human premalignant and malignant cutaneous disorders (Lippman & Meyskens, 1989). In addition, retinoids are successfully used in the therapy of the acute promyelocytic leukaemia (Meng-er et al., 1988).Several intracellular proteins interacting with RA have been identified, such as the cellular retinoic acid binding protein (CRABP) and the nuclear retinoic acid receptors (RARs). Three RAR subtypes RAR-a, -P and -y have been identified so far (Petkovich et al., 1987;Giguere et al., 1987;Benbrook et al., 1988;Brand et al., 1988;Krust et al., 1989 NIH:OVCAR-3 and the ovarian teratocarcinoma cell line CRL 1572 (PA-1) were received from the American Tissue Type Culture Collection (ATCC, Rockville, MD, USA). All cell lines were cultivated in a-MEM (Gibco, Scotland) supplemented with 10% heat-inactivated foetal calf serum (Gibco, Scotland) and were maintained in an humidified 5% CO2 atmosphere at 37°C. Cultures were refed after 4 days and passaged weekly 1:5 -1:10.All cells were used within 30 passages from the original stock.Tests for mycoplasma contamination were negative (DAPI, Boehringer Mannheim, Germany). RetinoidsAll-trans RA, 1 3-cis RA, (E)-4- [2-(5,6,7,8-tetrahydro-5,5,8,8,-tetramethyl-2-naphthalenyl)-1-propenyl] benzoic acid -(5,6,7,8-tetrahydro-5,5,8,8,-tetramethyl-2-

Growth-inhibitory effects of vitamin D analogues and retinoids on human pancreatic cancer cells

“…In contrast, we found differential expression of RAR-fl mRNA, which could be detected only in Hs766T cells, matching the product described previously (Harant et al, 1993) but neither in Capan 1 nor in Capan 2 cells (Figure 6). …”

“…weak signal of RAR-f3 and RAR-y (data not shown), we applied a seminested PCR as second amplification step for detection of the mRNA of these two receptors. Using this approach the three cell lines showed a strong signal of RARy mRNA (Figure 6) corresponding to the specific product described earlier in ovarian cancer cells (Harant et al, 1993).…”

“…Pancreatic cancer is poorly influenced by chemotherapy (Jeekel, 1994) (Reichel et al, 1989;Cross et al, 1992;Bollag and Holdener, 1992;Gudas, 1992 (Harant et al, 1993), breast cancer cell lines (Roman et al, 1992), myeloma cells and leukaemic cells (Lutzky et al, 1994;Dore et al, 1993). There is no correlation between biological effects of retinoids and degree of receptor expression (van-der Leede et al, 1993;Lutzky et al, 1994), which is consistent with our results.…”

Modulation of retinoic acid receptor function alters the growth inhibitory response of oral SCC cells to retinoids

“…Retinoids have been demonstrated to inhibit development of a number of di erent types of tumors (Aylsworth et al, 1986;Munker et al, 1987) and to inhibit the growth of many human tumor cell lines (Wu et al, 1997a(Wu et al, ,b, 1998aHarant et al, 1993;Saunders et al, 1993;Grunt et al, 1992;Somay et al, 1991Somay et al, , 1992Cline and Rice, 1983;Kim et al, 1984;Reiss et al, 1985;Thatcher et al, 1985;Rubin and Rice, 1986;Lotan et al, 1987;Sakar and Das, 1988) including squamous cell carcinomas (SCCs) (Cline and Rice, 1983;Kim et al, 1984;Reiss et al, 1985;Thatcher et al, 1985;Rubin and Rice, 1986;Lotan et al, 1987;Sakar and Das, 1988). Recently, retinoids have been successfully used in vivo to prevent the progression of preneoplastic oral, bronchial, and head-and-neck lesions to frank malignant tumors Benner et al, 1994;Lotan et al, 1995).…”

Retinoids: present role and future potential

“…Furthermore, tumour regression on treatment with retinoids has been demonstrated for in vivo xenograft models of experimental tumours including lip squamous cell carcinoma (Gottardis et al, 1996b) and melanoma (Schadendorf et al, 1996). Antiproliferative response appears to be mediated through the RARγ in melanoma and teratocarcinoma cell lines (Moasser et al, 1994;Schadendorf et al, 1994), although no correlation was noted with any receptor in ovarian cancer cell lines (Harant et al, 1993). Several groups have observed that differentiating agents such as butyrate and DMSO can induce p21 and terminal differentiation in a p53-independent manner, although p21 induction by differentiating agents can occur in the presence of wild-type p53 (reviewed in Liebermann et al, 1995).…”