Establishment and Characterization of a New Lung Cancer Cell Line (MI‐4) Producing High Levels of Granulocyte Colony Stimulating Factor

Abstract

Some cancer patients manifest leukocytosis without overt inflammation. The production of granulocyte colony-stimulating factor (G-CSF) by the tumor itself is thought to be responsible for this paraneoplastc syndrome.1-3) At least 80 cases of G-CSF-producing tumors have been reported up to the present. Although many types of G-CSF-producing tumors such as carcinoma of the thyroid, 4, 5) kidney, 6) oral cavity, 7) bladder, 8) gallbladder, 9) stomach, 10) hepatoma 11) and melanoma 12) have been reported, carcinoma of the lung [13][14][15][16][17] is predominant. Large cell carcinoma is the overwhelmingly dominant histologic type in lung cancer. 18,19) However, there have been few reports confirming the establishment of a G-CSF-producing cell line; to our knowledge, 10 G-CSF-producing lung cancer cell lines have been reported. Four of them are derived from large cell carcinoma. 14,[20][21][22] Recently, we studied a patient with large cell lung carcinoma who exhibited leukocytosis, and we established a new cell line in vitro from this patient. This cell line produces a high level of G-CSF in the medium. In this paper, we describe the establishment and characterization of this new lung cancer cell line. MATERIALS AND METHODS PatientThe patient was a 69-year-old male with large cell carcinoma of the lung. He underwent left upper lobectomy. Histologically, the surgical specimen of the lung tumor showed large cell undifferentiated carcinoma. However, the tumor metastasized to Virchow's node after 4 years. Chemotherapy and radiotherapy afforded only temporary control and the tumor grew rapidly. As the disease progressed, his peripheral blood white blood cell (WBC) count increased to 92 000/mm 3 with 95% mature neutrophils, without evidence of infection. The serum level of G-CSF was 141 pg/ml (normal <30 pg/ml). The patient gradually deteriorated with cachexia, and died 9 months after recurrence. At autopsy, metastatic tumors were found in the bilateral lungs, anterior mediastinum, right adrenal gland and retroperitoneal lymphnodes, besides Virchow's node. Furthermore, he had a large quantity of malignant pleural effusion and ascites. Cell culture Tumor cells were obtained from the pleural effusion and cultured for the establishment of a cell line.

The prognostic significance of leukocytosis in cervical cancer

“…Despite the fact that we had not investigated the presence in the serum and tumor of any of the cytokines, G‐CSF, GM‐CSF, IL‐6, IL‐1, and TNF cytokines have consistently been reported to be raised in dozens of reports dealing with paraneoplastic leukocytosis, and it has been firmly established that multiple hematopoietic cytokines particularly G‐CSF and GM‐CSF induce autocrine growth promotion in a number of cancer types and tumor models (19,29–35) . Further, TNF‐alpha and IL‐1beta enhance the expression of G‐CSF and GM‐CSF (36) . IL‐6 in particular has also been involved in paraneoplastic leukocytosis in cervical cancer.…”

Cyclooxygenase‐2 inhibitor NS‐398 suppresses cell growth and constitutive production of granulocyte‐colony stimulating factor and granulocyte macrophage‐colony stimulating factor in lung cancer cells

“…Several reports have demonstrated the presence of an autocrine growth loop for G-CSF 31,32) and GM-CSF 13) in non-hematopoietic tumor cells. However, our cell lines did not have an autocrine growth loop for G-CSF, 27) and they were not investigated for GM-CSF. To our knowledge, there have been no reports about autocrine growth control by G-CSF in lung cancer.…”

“…[38][39][40] We showed that the inflammatory cytokines, TNF-α and IL-1β induced COX-2 expression in addition to G-CSF or GM-CSF in OKa-C-1 and MI-4 cells. 27,41) Some studies have reported that NF-κB activa- tion is required to upregulate G-CSF, GM-CSF, or COX-2 expression, [42][43][44][45][46][47][48][49] and that inactivation of the NF-κB also induces rapid pro-apoptotic action. 45) COX-2 activation was also shown to follow the activation of MAPK in another cell type.…”

“…In our laboratory, we have established two lung cancer cell lines, OKa-C-1 and MI-4. 6,27) The OKa-C-1 and MI-4 lines were derived from patients with squamous cell carcinoma and large cell carcinoma, respectively. The two lung cancer cell lines have been maintained in RPMI 1640 medium containing 10% fetal calf serum (FCS) at 37°C in an incubator, under a 100% humidified 5% CO 2 atmosphere.…”

Establishment and characterization of human non-small cell lung cancer cell lines

“…3). In general, surgical tissues have been used to establish human lung cancer cell lines, and various functional lung cancer cell lines have been reported (10)(11)(12). Nonetheless, the incidence of lung cancer remains globally high.…”