Original Paper
Chromosomal alterations, biological features and in vitro chemosensitivity of SCLC-R1, a new cell line from human metastatic small cell lung carcinoma

https://doi.org/10.1016/S0959-8049(97)10148-4Get rights and content

Abstract

A new human cancer cell line was established from a metastatic lesion of a small cell lung carcinoma (SCLC-R1) and maintained in continuous culture with a doubling time of 62 h. The SCLC-R1 line, whose ultrastructural features are presented, showed a diploid DNA content, a translocation involving chromosome 16 [t(16;?)(q24;?)] and noticeable deletions in the FHIT (fragile histidine triad) region in the short arm of chromosome 3 [del(3)(p14)] and in the telomeric region of the short arm of chromosome 12 [del(12)(p13)]. The involvement of 12p in metastatic small cell lung cancer is reported here for the first time. No amplification or rearrangements were evident in the c-myc, L-myc, N-myc, int-2, c-erbB-2, H-ras, K-ras, c-mos, and hst-1 genes by Southern blot analysis. Wild-type p53, RB, K-ras and H-ras genes were evident by polymerase chain reaction–single-strand conformation polymorphism (PCR–SSCP) analysis. The neuron specific enolase (NSE) level was much higher in the cell line’s cytosol than in the patient’s serum and the cell line also had high expression of chromogranin A and cytokeratin 19. SCLC-R1 cells were sensitive to cisplatin, carboplatin and doxorubicin. The clinical history of the patient from whom the cell line was derived is reported. The characteristics of this new cell line indicate it to be a useful experimental model to investigate lung cancer biology and anticancer drug response.

Introduction

Lung cancer is the leading cause of cancer death, with mortality approaching 90% and a median survival of approximately 12 months[1]. Its incidence remains high in the industrialised world and is rapidly increasing in developing countries, due to the strong association with cigarette smoking[2].

The most common histological classification, proposed by the World Health Organization[3], divides lung tumours into four major categories, representing 95% of all lung cancers and among which 20–25% is due to small cell carcinoma. From a clinical point of view, bronchiogenic tumours are categorised into two groups, which reflect their biological behaviour and systemic or local therapeutic management: small cell lung cancers (SCLC) and non-small cell lung cancers (NSCLC), including adenocarcinomas, squamous cell and large cell carcinomas. The interface between the two groups needs frequent revision due to the advances in knowledge of their cellular and molecular biology[4].

It is known that both inherited predisposition and acquired somatic genetic changes contribute to lung tumour development and progression[5]. It has been postulated that such genetic events may be related to the resistance to chemotherapy, either intrinsic or acquired, which represents one of the major obstacles preventing cure in patients affected by SCLC[6]. The availability of well characterised lung tumour cell lines, derived directly from patients with a documented clinical history and retaining properties observed in vivo, provides fundamental models for genetic and biological studies and for in vitro search of more effective treatments[7].

We report here the establishment and characterisation of a new cell line derived from a metastatic small cell carcinoma of the lung (SCLC-R1). The clinical history of the patient from whom the cells were derived is presented, together with the ultrastructural, cytogenetic and molecular features of the cells. The results of tumour markers, immunohistochemistry and chemosensitivity assays are shown in comparison with those evident in the patient.

Section snippets

Clinical findings

The cell line was obtained from a metastatic supraclavear lymph node of a 65 year old man affected by SCLC (stage III). The patient had been treated with endoxan, 4′-epidoxorubicin and etoposide, with a response higher than 80%. Surgical resection of the lung was then performed. Three months later, metastatic supraclavear lymph nodes were detected (cell line derived from one of these) and a new treatment (cisplatin plus etoposide) was started. Eight months later the patient died of

Results

The SCLC-R1 cell line was derived from one tumour sample, grown free of mycoplasma contamination, mostly as a monolayer, and had a doubling time of approximately 62 h. By phase contrast, the cells appeared prevailingly spindle shaped and grew several layers deep with no contact inhibition. Some superficial cells tended to detach (data not shown). By electron microscopy, in sections parallel to the culture surface, the cells were flattened and spindle shaped and the cell membrane was smooth, with

Discussion

Preventive care measures, earlier diagnosis and more effective treatments are the main research objectives to reduce both the incidence and the mortality rates of lung cancer. For these purposes and to understand better lung tumorigenesis and progression, the availability of well characterised tumour cell lines derived directly from patients with a documented clinical history and retaining properties observed in vivo is important. We report here the characteristics of a new cell line derived

Acknowledgements

This work was supported by grants from the Istituto Oncologico Romagnolo Forli, the A.I.R.C. Milano, the C.N.R.-ACRO Project (no. 920227/3-PF 39), the MURST 40%–60% and the University of Bologna (Funds for selected topics), Italy. The probes for fluorescent in situ hybridisation analysis were a generous gift from Dr Peter Marynen, Centre for Human Genetics, Leuven, Belgium. Docetaxel was a kind gift from Rhône-Poulenc Rorer.

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