Saleem Malik

The detection and localization of monocyte chemoattractant protein-1 (MCP-1) in human ovarian cancer.

Chemokines may control the macrophage infiltrate found in many solid tumors. In human ovarian cancer, in situ hybridization detected mRNA for the macrophage chemokine monocyte chemoattractant protein-1 (MCP-1) in 16/17 serous carcinomas, 4/4 mucinous carcinomas, 2/2 endometrioid carcinomas, and 1/3 borderline tumors. In serous tumors, mRNA expression mainly localized to the epithelial areas, as did immunoreactive MCP-1 protein. In the other tumors, both stromal and epithelial expression were seen. All tumors contained variable numbers of cells positive for the macrophage marker CD68. MCP-1 mRNA was also detected in the stroma of 5/5 normal ovaries. RT-PCR demonstrated mRNA for MCP-1 in 7/7 serous carcinomas and 6/6 ovarian cancer cell lines. MCP-1 protein was detected by ELISA in ascites from patients with ovarian cancer (mean 4.28 ng/ml) and was produced primarily by the cancer cells. Human MCP-1 protein was also detected in culture supernatants from cell lines and in ascites from human ovarian tumor xenografts which induce a peritoneal monocytosis in nude mice. We conclude that the macrophage chemoattractant MCP-1 is produced by epithelial ovarian cancer and that the tumor cells themselves are probably a major source. MCP-1 may contribute to the accumulation of tumor-associated macrophages, which may subsequently influence tumor behavior.

Cells Secreting Tumour Necrosis Factor Show Enhanced Metastasis in Nude Mice

A tumour cell may acquire the ability to invade and metastasise via heritable changes in its genome and/or changes in the local environment. Chinese hamster ovary (CHO) cells transfected with the gene for human TNF (CHO/TNF cells) showed a greatly enhanced ability to invade peritoneal surfaces and metastasise in nude mice compared with cells transfected by the vector alone. In situ hybridisation with a riboprobe for human TNF showed that the CHO/TNF cells were actively transcribing this cytokine after in vivo injection. Neutralising antibodies to human TNF, both whole IgG and F(ab)2 fragments, abrogated the enhanced metastatic activity of the TNF-secreting cells. Thus transfection of a cytokine/growth-factor gene can confer a metastatic phenotype on the recipient cell.

In situ detection of tumour necrosis factor in human ovarian cancer specimens.

In situ hybridisation was used to study the local expression of tumour necrosis factor (TNF) mRNA in human ovarian tumours. In 8 of 14 ovarian cancers studied, a minority of cells in the epithelial areas of the tumour contained TNF mRNA. In individual high-power fields as many as 8% of cells were positive for TNF mRNA. Immunohistochemical studies on sequential sections and the morphology of the positive cells led to the conclusion that the ovarian tumour cells were transcribing the TNF gene. There was immunohistochemical evidence of the production of TNF protein by the tumour cells and TNF protein in a tumour lysate. The production of TNF by human ovarian cancer cells may influence the biology of the tumour, contribute to neoplastic progression and alter the response to therapy.

Tumour necrosis factor as an anticancer agent

WHEN the human gene encoding tumour necrosis factor (TNF) was cloned in 1984 [ 11, there was excitement at the prospect of large amounts of purified recombinant material for clinical cancer trials. This factor was potentially an important advance in cancer treatment. The historical background [2] and preclinical studies with partially purified material [3] gave grounds for cautious optimism. Manda et al. [4] reported on the antitumour effect of TNF and S-fluorouracil in the murine Meth A sarcoma model, and highlighted the importance of tumour blood supply to the antitumour effect of TNF. This adds to the large body of data now available in animal models on the antitumour efficacy of TNF as a single agent, or in combination with other cytokines and cytotoxic drugs. However, the promising results in animal models have not, as yet, been reflected in clinical trial data. Six years after the TNF gene was cloned there is little evidence for antitumour activity of this agent in man, although most of the completed studies are phase I trials [5]. An understanding of the history of TNF and the advances made since the gene was cloned provides some explanation for the disappointing results obtained in clinical cancer trials. Such information may indicate ways in which the undoubted power of this cytokine could be effectively used singly or in combination with other antitumour agents. In addition, there is now evidence that endogenous TNF production may be inextricably bound with growth and metastasis of some tumours, and that neutralization of TNF activity may be of potential therapeutic benefit.

Tumour necrosis factor.

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Therapy of human ovarian cancer xenografts with intraperitoneal liposome encapsulated muramyl-tripeptide phosphoethanolamine (MTP-PE) and recombinant GM-CSF

Three intraperitoneal human ovarian cancer xenografts (OS, HU, and LA) were used to assess the antitumour activity of intraperitoneal therapy with liposome encapsulated MTP-PE. MTP-PE led to significant prolongation of survival in all three xenograft models, but with varying efficacy. In one tumour model (OS), 80% of mice were cured of tumour by twice weekly therapy for 4 weeks, whereas in another xenograft model (LA), the median survival time was approximately doubled compared to PBS injected and placebo liposome injected controls (median survivals: 30 vs 62.5 days respectively). The antitumor efficacy of MTP-PE did not correlate with the extent of peritoneal neutrophil infiltration after intraperitoneal therapy. Combined therapy with liposome encapsulated MTP-PE and recombinant murine granulocyte-macrophage colony stimulating factor led to increased survival of mice bearing the LA and HU xenografts, compared to tumour bearing mice treated with either agent singly.

Effects of intraperitoneal recombinant interleukin-1 beta in intraperitoneal human ovarian cancer xenograft models: comparison with the effects of tumour necrosis factor.

The effect of intraperitoneal (i.p.) injection of recombinant human interleukin-1 beta (rhIL-1 beta) was studied in three i.p. nude mouse xenograft models of human ovarian cancer (HU, OS, and LA). Intraperitoneal rhIL-1 beta administration led to a dose dependent replacement of peritoneal ascitic tumour with solid tumours attached to the peritoneum and intraabdominal viscera in two (HU and LA) out of the three xenograft models. In the third xenograft model (OS), low doses of rhIL-1 beta (10 ng day) promoted micrometastatic peritoneal implants of tumour, but higher doses of rhIL-1 beta (1 microgram day) had a marked antitumour effect. This was due to direct cytotoxicity for tumour cells and was not related to peritoneal neutrophil influx induced by rhIL-1 beta. Recombinant human TNF (rhTNF) also promoted tumour implantation in all three xenograft models, but its antitumour effects differed from rhIL-1 beta. TNF increased the survival of HU and LA bearing mice, but had no antitumour effect in the OS xenograft model. Analysis of peritoneal fluid and tumour xenografts showed that TNF induced murine IL-1 in the tumour bearing mice. The magnitude of IL-1 induction indicated that TNF induced IL-1 did not contribute significantly to its effects.