Caspase-3 activity predicts local recurrence in rectal cancer.

PURPOSE: Radiotherapy followed by total mesorectal excision surgery has been shown to significantly reduce local recurrence rates in rectal cancer patients. Radiotherapy, however, is associated with considerable morbidity. The present study evaluated the use of biochemical detection of enzymatic caspase-3 activity as preoperative marker for apoptosis to preselect patients that are unlikely to develop a local recurrence to spare these patients from overtreatment and the negative side effects of radiotherapy. EXPERIMENTAL DESIGN: Nonirradiated freshly frozen tissue samples from 117 stage III rectal cancer patients were collected from a randomized clinical trial that evaluated preoperative radiotherapy in total mesorectal excision surgery. Additional frozen archival tissues from 47 preoperative biopsies and corresponding resected colorectal tumors were collected. Level of apoptosis was determined by measuring the enzymatic activity of caspase-3 in a biochemical assay. RESULTS: In tumor tissue, caspase-3 activity lower than the median was predictive of 5-year local recurrence (hazard ratio, 7.4; 95% confidence interval, 1.7-32.8; P = 0.008), which was unaffected by adjustment for type of resection, tumor location, and T status (adjusted hazard ratio, 7.5; 95% confidence interval, 1.7-34.1; P = 0.009). Caspase-3 activity in preoperative biopsies was significantly correlated with caspase-3 activity in corresponding resected tumors (r = 0.56; P < 0.0001). CONCLUSION: Detection of tumor apoptosis levels by measuring caspase-3 activity, for which a preoperative biopsy can be used, accurately predicted local recurrence in rectal cancer patients. These findings indicate that caspase-3 activity is an important denominator of local recurrence and should be evaluated prospectively to be added to the criteria to select rectal cancer patients in which radiotherapy is redundant.

A systemic antitumor immune response prevents outgrowth of lung tumors after i.v. rechallenge but is not able to prevent growth of experimental liver tumors.

Although a systemic antitumor immune response by antibodies or T cells is often detected in cancer patients, this response mostly does not result in tumor rejection. The beneficial effect of tumor vaccination on survival rates is limited as tumor response is low. In contrast to solid tumors, circulating tumor cells may be more easily accessible and therefore destroyed by the immune system and thus prevent metastases. This discrepancy is further clarified in our study, by assessing the effect of a systemic immune response on established liver tumors and circulating tumor cells. Male Wag/Rij rats were inoculated with CC531 colorectal tumor cells subcapsulary in the liver, with or without immune suppression (60 mg/kg cyclophosphamide). To study the effect of a systemic immune response, rats received CC531 tumor cells intravenously and three weeks later the number of lung tumors was assessed. Presence of specific anti-CC531 antibodies in serum was determined by flow cytometric analysis at times of inoculation, i.v. tumor cell administration and sacrifice. Rats with liver tumors and subsequent rechallenge produced anti-CC531 IgG antibodies and did not develop lung tumors, whereas without existing liver tumors, rats developed lung tumors upon i.v. administration of CC531 tumor cells. Liver tumors in rats with and without i.v. CC531 tumor cell administration were equal in size. These results showed that a systemic immune response, induced upon liver tumor induction and rechallenge, prevented formation of lung tumors but did not affect tumor growth in the liver. Possibly the immune response lacked the ability to penetrate the protective extracellular matrix surrounding the established liver tumors, which prevented the tumor cells from recognition by and contact with cells of the immune system.

The immunological consequences of photodynamic treatment of cancer, a literature review.

In this review we discuss the effect of photodynamic treatment (PDT) of solid tumors on the immune response. The effect on both the innate and adapted immune response is discussed. We have summarized the evidence that PDT causes or enhances an anti-tumor response. PDT is a local treatment in which the treated tumor remains in situ while the immune system is only locally affected and still functional in contrast with e.g. after systemic chemotherapy. We conclude that PDT of cancer is a way of in situ vaccination to induce a systemic antitumor response. In general, immune cells are found in the tumor stroma, separated from tumor cells by extracellular matrix and basal membrane-like structures. We hypothesize that PDT destroys the structure of a tumor, thereby enabling direct interaction between immune cells and tumor cells resulting in the systemic anti-tumor immune response.

Rat colon carcinoma cells that survived systemic immune surveillance are less sensitive to NK-cell mediated apoptosis.

In order to form distant metastases, cells from the primary tumor have to detach, enter the blood- or lymph-compartment and escape immune surveillance. Here, we describe the selection of rat colon carcinoma cell lines (CC531s-m1 and CC531s-m2) that escaped from systemic immune surveillance; CC531s cells were injected into the v. jugularis of Wag/Rij rats, after three weeks the lung tumors were isolated, the tumor cells were cultured, characterized and injected again. The m1- and m2-cell lines were less susceptible for killing by syngeneic NK cells. Further characterization of this cell line showed a decreased sensitivity towards TRAIL- and CD95L-, but not to granzyme B-mediated apoptosis. In the m1- and m2-cells log-phase growth started earlier as compared to the parental cell line, whereas no changes were found in anchorage-dependent or anchorage-independent growth. After subcapsular injection of the m2-cell line into the liver of rats much more lung metastases were formed in comparison to injection of the parental cell line. In conclusion, the results suggest that the resistance of the m1- and m2-cells to NK cell-mediated apoptosis was associated with their capability to survive systemic immune surveillance and form metastases in vivo.

A tumor-specific kinase activity regulates the viral death protein Apoptin.

Apoptin, a chicken anemia virus-encoded protein, is thought to be activated by a general tumor-specific pathway, because it induces apoptosis in a large number of human tumor or transformed cells but not in their normal, healthy counterparts. Here, we show that Apoptin is phosphorylated robustly both in vitro and in vivo in tumor cells but negligibly in normal cells, and we map the site to threonine 108. A gain-of-function point mutation (T108E) conferred upon Apoptin the ability to accumulate in the nucleus and kill normal cells, implying that phosphorylation is a key regulator of the tumor-specific properties of Apoptin. An activity that could phosphorylate Apoptin on threonine 108 was found specifically in tumor and transformed cells from a variety of tissue origins, suggesting that activation of this kinase is generally associated with the cancerous or pre-cancerous state. Moreover, analyses of human tissue samples confirm that Apoptin kinase activity is detectable in primary malignancies but not in tissue derived from healthy individuals. Taken together, our results support a model whereby the dysregulation of the cellular pathway leading to the phosphorylation of Apoptin contributes to human tumorigenesis.