Phenotypic characterization, karyotype analysis and in vitro tamoxifen sensitivity of new ER‐negative vulvar carcinoma cell lines, UM‐SCV‐1A and UM‐SCV‐1B

Abstract

Two cell lines (University of Michigan squamous carcinoma of the vulva UM-SCV-1A and UM-SCV-1B) were established from the primary tumor and a malignant pleural effusion of a 62-year-old woman. Both tumor specimens grew vigorously in vitro and could be passaged after only 14 and 10 days in culture, respectively. Both cell lines undergo 3 population doublings in 4 days, reaching saturation densities of 5 x 10(5) cells/cm2, and have been carried through more than 30 in vitro passages. In nude mice the cultured cells initially formed tumors but these regressed 2-3 weeks after inoculation. The regressing mouse tumors consisted of poorly differentiated squamous carcinoma surrounded by an inflammatory lymphoid infiltrate. The UM-SCV-1 cell lines express membrane antigens typically displayed by squamous-cell carcinomas. These include the HLA class-1 light chain beta 2-microglobulin, pemphigus, pemphigoid, and the alpha 6 beta 4 integrin defined by the UM-A9 monoclonal antibody (MAb). In contrast to the A431 vulvar carcinoma, these tumor lines do not have amplified expression of the epidermal growth factor (EGF) receptor. Although tissue from the primary tumor contained low levels of estrogen receptor activity, no receptor activity was detected in the cell lines. Nevertheless, both lines were sensitive to growth inhibition by tamoxifen. This effect was not reversible by estradiol, indicating an estrogen-receptor-independent mechanism. The tumors were both hypotetraploid, contained the same chromosome rearrangements and had stable karyotypes in vitro. Each contained inv(1)(p36.3q32.1), del(4)(q12), dic(4;11)(q12;p11.2), i(5p), der(6)t(3;6)(q25.1;p21.1), several rearrangements involving chromosomes 8 and 14, + i(13), i(18p), a dicentric t(11;19), and 2 or 3 unidentified markers. Since the karyotypes of both tumors were the same, no major karyotypic change was associated with metastatic spread. These paired primary and metastatic SCC lines from an unusually aggressive vulvar carcinoma provide an in vitro model for analysis of the biological basis of this tumor's behavior.

Paclitaxel combined with fractionated radiation in vitro: A study with vulvar squamous cell carcinoma cell lines

“…24,25 The longestablished vulvar SCC cell line A-431 was obtained from the American Type Culture Collection (Rockville, MD). 26 Some characteristics of the cell lines are listed in Table I.…”

Identification of individual genes altered in squamous cell carcinoma of the vulva

“…Data on patient age, tumor-node-metastasis (TNM) disease classification, histologic grade of the tumor, prior therapy at the time of tissue biopsy for establishment of the cell line, and site of the specimen have been previously published (Gré nman et al, 1990;Worsham et al, 1991;Raitanen et al, 2004). Cell cultures were started from fresh tumor samples and have been studied in detail (Gré nman et al, 1990;Worsham et al, 1991;Raitanen et al, 2004) for cell growth characteristics, chromosome analysis, CGH, growth in nude mice, flow cytometry, expression of cytokeratin, and mycoplasma status.…”

“…Cell cultures were started from fresh tumor samples and have been studied in detail (Gré nman et al, 1990;Worsham et al, 1991;Raitanen et al, 2004) for cell growth characteristics, chromosome analysis, CGH, growth in nude mice, flow cytometry, expression of cytokeratin, and mycoplasma status.…”

“…DNA, extracted using the QIAamp Kit (Qiagen Inc, Chatsworth, CA), was obtained from passaged cell lines (Table 1) that originated from fresh tumor samples as described previously (Carey, 1985;Gré nman et al, 1988Gré nman et al, , 1990. DNA was extracted from each cell line at a specific passage number (P) as follows: UM-SCV-1A, P22; UM-SCV-1B, P8; UM-SCV-2, P22; UM-SCV-3, P26; UM-SCV-4, P19; UM-SCV-6, P17; UM-SCV-7, P13; UT-SCV-1, P26: UT-SCV-2 and -3, P27; UT-SCV-4, P12; UT-SCV-5, P9; and UT-SCV-6, P12.…”

Retargeting improves the efficacy of a telomerase-dependent oncolytic adenovirus for head and neck cancer

“…The head and neck squamous cell carcinoma cell lines (UT-SCCs) used in this study were obtained from Turku University Central Hospital, where these cell lines were established during surgery from primary or metastasized tumors (29). The cells were cultured at 37˚C with 5% CO 2 in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, non-essential amino acids and penicillin/streptomycin.…”