Chemotherapy For Melanoma Research Articles

O6-methylguanine DNA-methyltransferase (MGMT) overexpression in melanoma cells induces resistance to nitrosoureas and temozolomide but sensitizes to mitomycin C

Alkylating agents play an important role in the chemotherapy of malignant melanomas. The activity of alkylating agents depends on their capacity to form alkyl adducts with DNA, in some cases causing cross-linking of DNA strands. However, the use of these agents is limited by cellular resistance induced by the DNA repair enzyme O(6)-methylguanine DNA-methyltransferase (MGMT) which removes alkyl groups from alkylated DNA strands. To determine to what extent the expression of MGMT in melanoma cells induces resistance to alkylating agents, the human cell line CAL77 Mer- (i.e., MGMT deficient) were transfected with pcMGMT vector containing human MGMT cDNA. Several clones expressing MGMT at a high level were selected to determine their sensitivity to chemotherapeutic drugs. Melanoma-transfected cells were found to be significantly less sensitive to nitrosoureas (carmustine, fotemustine, streptozotocin) and temozolomide with an increase of IC(50) values between 3 and 14 when compared to parent cells. No difference in cell survival rates between MGMT-proficient and -deficient cells was observed for melphalan, chlorambucil, busulphan, thiotepa and cisplatin which preferentially induce N(7) guanine lesions. Surprisingly, MGMT overexpression increased the sensitivity of CAL77 cells to mitomycin C by approximately 10-fold. Treatment of clonal cell lines with buthionine-[S,R]-sulfoximine (BSO), an inhibitor of gamma-glutamylcysteine synthetase which depletes cellular glutathione, completely reversed this unexpected increase in sensitivity to mitomycin C. This observation suggests that glutathione is involved in the sensitivity of MGMT-transfected cells to mitomycin C and may act synergistically with MGMT via an unknown mechanism.

Combined chemoimmunotherapy of metastatic melanoma: a single institution experience

The addition of cytokines, such as interferon alpha-2b and interleukin-2, to chemotherapy in metastatic melanoma has produced conflicting results in phase II and III trials. We report our experience with a chemoimmunotherapeutic regimen using subcutaneous cytokines. Twenty-eight patients with advanced melanoma (median age, 45 years; male to female ratio, 19 : 9) were treated. Doses were as follows: cisplatin, 20 mg/m intravenously (iv) days 1-4; vinblastine, 1.6 mg/m iv days 1-4; dacarbazine, 800 mg/m iv day 1; interferon alpha-2b, 5 MIU/m subcutaneously (sc) days 1-5; interleukin-2, 9 MIU/m sc days 1-5 and 8-12. Treatment was repeated every 3 weeks for a maximum of six cycles. The response was assessed after two cycles and toxicity at every cycle, according to World Health Organization (WHO) and National Cancer Institute (NCI) criteria, respectively. At a median follow-up of 8 months, only four patients (14%) were still alive. The overall response rate was 33%, with three (11%) complete responses lasting for 17, 14 and >24 months. There were six (22%) partial responses and three stable disease. Amongst the responders, three patients progressed at the level of the central nervous system. The median time to progression and overall survival were 3.5 and 9 months, respectively. The most common grade 3-4 toxicity was neutropenia, reported in 25 of the 28 patients (92%). Only two patients (7%) experienced neutropenic fever. Thrombocytopenia grade 3-4 occurred in seven of the 28 patients (25%), with only one patient needing transfusional support. One toxic death due to neutropenic fever occurred. It can be concluded that the chemoimmunotherapy schedule evaluated is active and may be considered for patients with metastatic melanoma who have a good performance status and a limited disease burden.

Hughes Technique, Amniotic Membrane Allograft, and Topical Chemotherapy in Conjunctival Melanoma With Eyelid Involvement

A 59-year-old woman presented with a pigmented mass in the inferior tarsal conjunctiva of the left eye with an associated diffuse, multifocal pigmentation involving largely the inferior half of the bulbar conjunctiva, fornix, and eyelid skin. Histopathologic examination of map biopsies disclosed conjunctival melanoma from primary acquired melanosis. Surgical excision of the inferior bulbar conjunctiva, fornix, and lower eyelid with histopathologic free margins was performed. Adjuvant cryotherapy was applied. The bulbar conjunctiva and lower fornix were reconstructed with an amniotic membrane allograft. Lower eyelid reconstruction was accomplished by use of the Hughes technique. Topical mitomycin C (0.04%) was applied after surgery. After 2 years of follow-up, no tumor recurrence has been detected and the eyelid and conjunctival defect have been satisfactorily corrected. This combined surgical procedure using amniotic membrane allograft and a composite tarsoconjunctival flap is shown to be useful in the treatment of an advanced conjunctival neoplasia with extensive eyelid involvement.

Antitumor Efficacy of bcl-2 and c-myc Antisense Oligonucleotides in Combination with Cisplatin in Human Melanoma Xenografts: Relevance of the Administration Sequence

bcl-2 and c-myc oncogenes are frequently overexpressed in different human tumors, including melanoma. Here, we evaluate the combined efficacy of two antisense oligonucleotides targeting bcl-2 mRNA (ODN bcl-2) and c-myc mRNA (ODN c-myc) in combination with cis-diammine dichloroplatinum (cisplatin, DDP) on three human melanoma lines (LM, NG, and M20). Two different sequences were designed to treat tumor-bearing mice: in the first one, ODN bcl-2 at a dose of 0.2 mg/day x4, followed by DDP given i.p. at a dose of 3.3 mg/kg/day x3 and ODN c-myc i.v. at 0.5 mg/day x7, whereas the other sequence consisted of ODN c-myc given as first agent followed by DDP and ODN bcl-2 at the same doses. Mice received three complete cycles of treatment in 1-week intervals. The treatment sequence with ODN bcl-2/DDP/ODN c-myc combination completely inhibited growth in NG tumor and induced a 35-day delay in LM tumor growth. In contrast, the M20 tumor growth was unaffected by the combination. A discrete amount of c-Myc and bcl-2 protein expression in both LM and NG tumors was detected, whereas no detectable levels of the two proteins were observed in M20 tumors. Compared with the other combination, the sequence (ODN bcl-2/DDP/ODN c-myc) produced the most effective results, producing a significant decrease in bcl-2 and c-Myc protein expression, which in turn significantly increased the survival of NG- and LM-bearing mice, with 4 mice out of 11 and 1 out of 7 mice being cured, respectively. Finally, this combination increased the apoptotic rate and produced an antiangiogenetic effect. These results show that an antisense approach to the treatment of melanoma xenografts overexpressing either bcl-2 or c-myc oncogenes represents a successful strategy to improve the response to chemotherapy in melanoma, with particular attention to the treatment sequence.

Psoralen derivatives and longwave ultraviolet irradiation are active in vitro against human melanoma cell line

Cutaneous malignant melanoma is a very serious form of skin cancer that arises from melanocytes. Currently there is no effective treatment for metastatic melanoma so intense clinical trials are evaluating new drugs for this human malignancy. Psoralens are a group of compounds that bind to DNA in rapidly dividing cells and with ultraviolet light in the A band (UVA) cause DNA crosslinking, thereby preventing cellular division. They are used in the treatment of psoriasis and cutaneous T-cell lymphoma among other skin and blood diseases. We have investigated the cytotoxic potential of three psoralen derivatives plus UVA exposure (PUVA) on a established cell line of human melanoma. Cells were treated with different concentrations of 8-methoxypsoralen (8-MOP), 4,5′,8-trimethylpsoralen (TMP) and 7-methylpyridopsoralen (MPP), for 1 h and after exposure to UVA light (0.3 J/cm 2 ) were allowed to recover over a 24–72 h period. Viability was assessed by the microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Cisplatin, one of the most important drugs in the chemotherapy of melanoma, was included for comparative studies. All the psoralen derivatives tested were markedly cytotoxic in a dose and post-exposure-time dependent manner. The IC 50 values for 72 h of post-exposure time were as follows: MPP = 0.05 ± 0.01, TMP = 0.13 ± 0.003 and 8-MOP = 10.79 ± 1.85 μmol/L. Regardless of the limitations of the in vitro model, our results suggested that the lower IC 50 values of TMP and MPP might be of clinical importance.

Psoralen derivatives and longwave ultraviolet irradiation are active in vitro against human melanoma cell line

Cutaneous malignant melanoma is a very serious form of skin cancer that arises from melanocytes. Currently there is no effective treatment for metastatic melanoma so intense clinical trials are evaluating new drugs for this human malignancy. Psoralens are a group of compounds that bind to DNA in rapidly dividing cells and with ultraviolet light in the A band (UVA) cause DNA crosslinking, thereby preventing cellular division. They are used in the treatment of psoriasis and cutaneous T-cell lymphoma among other skin and blood diseases. We have investigated the cytotoxic potential of three psoralen derivatives plus UVA exposure (PUVA) on a established cell line of human melanoma. Cells were treated with different concentrations of 8-methoxypsoralen (8-MOP), 4,5′,8-trimethylpsoralen (TMP) and 7-methylpyridopsoralen (MPP), for 1 h and after exposure to UVA light (0.3 J/cm 2) were allowed to recover over a 24–72 h period. Viability was assessed by the microculture 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric assay. Cisplatin, one of the most important drugs in the chemotherapy of melanoma, was included for comparative studies. All the psoralen derivatives tested were markedly cytotoxic in a dose and post-exposure-time dependent manner. The IC 50 values for 72 h of post-exposure time were as follows: MPP = 0.05 ± 0.01, TMP = 0.13 ± 0.003 and 8-MOP = 10.79 ± 1.85 μmol/L. Regardless of the limitations of the in vitro model, our results suggested that the lower IC 50 values of TMP and MPP might be of clinical importance.

Dacarbacine (DTIC) versus vaccination with autologous peptide-pulsed dendritic cells (DC) as first-line treatment of patients with metastatic melanoma: Results of a prospective-randomized phase III study

7508 Background: Progress in the treatment of patients with metastatic melanoma has been very limited over the last two decades. Vaccination with autologous DC was reported as being able to induce some clinical regression in early clinical phase I/II studies. Methods: Patients with advanced metastatic melanoma, HLA-A1, -A2, -A3, -A24 or -B44 positivity, no brain or bone metastases and any previous systemic chemotherapy were randomised to receive DTIC (850 mg/m2 d1, every 28 days with 4-week intervals (arm A) or intradermal vaccination with autologous peptide-pulsed dendritic cells in 2-week intervals for the first 5 vaccinations and every 4 weeks thereafter (arm B). DC were generated from peripheral blood after leukapheresis and culturing monocytes in-vitro in GM-CSF, IL-4 (d1-d5) and a maturing cytokine cocktail (IL-1β, IL-6, TNFα and PGE2) on day 6. Matured DC were pulsed subsequently with peptides derived from MAGE-1, MAGE-3, Melan-A, gp100, tyrosinase and influenza proteins as a control depending on the individual HLA-type (d6). Primary study endpoint was objective response to treatment (OR), secondary endpoints included safety, overall and progression-free survival. The planned accrual per arm was set at 119 patients under the basic assumption that the response rate would double from 15 % for treatment with DTIC to 30 % for DC-vaccination to reach significance at 0.05 with 80% power. Results: In the time period between 1/2001 and 6/2003 a total of 108 (intention-to-treat population, ITT; 55 DTIC, 53 DC) patients were randomized from 1 Swiss and 5 German centers with experience in DC generation and therapy. Accrual was halted prematurely by the data monitoring board because data indicated that the study goals could not be achieved. OR (CR+PR) were 5.5% in arm A and 3.8% in arm B, respectively (ITT). There was no statistically significant difference for response, toxicity, overall and progression-free survival between both study arms. Subset analysis indicated a trend for benefical effect of HLA-A2 and a disadvantage for HLA-B44. Conclusion: DC-based vaccination is as poorly active as standard DTIC chemotherapy in metastatic melanoma. Whether further selection of patients and improved DC generation protocols will lead to a statistical benefit remains topic of further studies. Author Disclosure Employment or Leadership Consultant or Advisory Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Merix

Dacarbacine (DTIC) versus vaccination with autologous peptide-pulsed dendritic cells (DC) as first-line treatment of patients with metastatic melanoma: Results of a prospective-randomized phase III study

7508 Background: Progress in the treatment of patients with metastatic melanoma has been very limited over the last two decades. Vaccination with autologous DC was reported as being able to induce some clinical regression in early clinical phase I/II studies. Methods: Patients with advanced metastatic melanoma, HLA-A1, -A2, -A3, -A24 or -B44 positivity, no brain or bone metastases and any previous systemic chemotherapy were randomised to receive DTIC (850 mg/m2 d1, every 28 days with 4-week intervals (arm A) or intradermal vaccination with autologous peptide-pulsed dendritic cells in 2-week intervals for the first 5 vaccinations and every 4 weeks thereafter (arm B). DC were generated from peripheral blood after leukapheresis and culturing monocytes in-vitro in GM-CSF, IL-4 (d1-d5) and a maturing cytokine cocktail (IL-1β, IL-6, TNFα and PGE2) on day 6. Matured DC were pulsed subsequently with peptides derived from MAGE-1, MAGE-3, Melan-A, gp100, tyrosinase and influenza proteins as a control depending on the individual HLA-type (d6). Primary study endpoint was objective response to treatment (OR), secondary endpoints included safety, overall and progression-free survival. The planned accrual per arm was set at 119 patients under the basic assumption that the response rate would double from 15 % for treatment with DTIC to 30 % for DC-vaccination to reach significance at 0.05 with 80% power. Results: In the time period between 1/2001 and 6/2003 a total of 108 (intention-to-treat population, ITT; 55 DTIC, 53 DC) patients were randomized from 1 Swiss and 5 German centers with experience in DC generation and therapy. Accrual was halted prematurely by the data monitoring board because data indicated that the study goals could not be achieved. OR (CR+PR) were 5.5% in arm A and 3.8% in arm B, respectively (ITT). There was no statistically significant difference for response, toxicity, overall and progression-free survival between both study arms. Subset analysis indicated a trend for benefical effect of HLA-A2 and a disadvantage for HLA-B44. Conclusion: DC-based vaccination is as poorly active as standard DTIC chemotherapy in metastatic melanoma. Whether further selection of patients and improved DC generation protocols will lead to a statistical benefit remains topic of further studies. Author Disclosure Employment or Leadership Consultant or Advisory Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Merix

Vinorelbine in Combination with Interleukin-2 as Second-Line Treatment in Patients with Metastatic Melanoma. A Phase II Study of the Hellenic Cooperative Oncology Group

Objectives: To evaluate the efficacy and toxicity of the combination of vinorelbine and interleukin (IL)-2 in patients with metastatic melanoma as second-line chemotherapy. Patients and Methods: Twenty-two patients with histologically confirmed stage IV melanoma previously treated with temozolomide-based chemotherapy—only one regimen of chemotherapy for disseminated disease was allowed—were treated with vinorelbine 30 mg/m2 on days 1 and 15 and IL-2 subcutaneous 9 × 106 once daily on days 2–6 and 16–19 every 4 weeks for maximum of six cycles. Results: From January 2000 to July 2001, 22 patients entered the study; the median age was 56 years. Among 20 evaluable patients there were 2 (9.1%) objective responses including 1 complete response and 1 partial response. Five (22.7%) had stabilization of their disease, and 13 (59.1%) progressed. The median time to progression (TTP) was 2.9 months and the median overall survival was 9.1 months. There was a significant difference in TTP in patients who responded or remained stable (median TTP 10.75 months) and those who progressed (median TTP 2.1 months) (p < 0.05). There was also a difference in survival in the two groups (p < 0.05 (28 vs. 8 months). The most common side effects were flulike symptoms, such as fever, chills, fatigue, and injection site reaction. Grade 3 hematological toxicity rarely occurred. One patient discontinued therapy because of fatigue and anorexia. There were no treatment-related deaths. Conclusions: The combination of vinorelbine and IL-2 provides clinical benefit in patients recurring or progressing on first-line chemotherapy for metastatic melanoma, with manageable toxicity.

Relation between RNA expression of DNA repair genes and ifosfamide chemotherapy of melanomas in vitro

Relation between RNA expression of DNA repair genes and ifosfamide chemotherapy of melanomas in vitro

834 Second-line treatment of disseminated malignant melanoma with Fotemustine

Alkylating agents play an important role in the chemotherapy of malignant melanomas. The activity of alkylating agents depends on their capacity to form alkyl adducts with DNA, in some cases causing cross-linking of DNA strands. However, the use of these agents is limited by cellular resistance induced by the DNA repair enzyme O6-methylguanine DNA-methyltransferase (MGMT) which removes alkyl groups from alkylated DNA strands. To determine to what extent the expression of MGMT in melanoma cells induces resistance to alkylating agents, the human cell line CAL77 Mer− (i.e., MGMT deficient) were transfected with pcMGMT vector containing human MGMT cDNA. Several clones expressing MGMT at a high level were selected to determine their sensitivity to chemotherapeutic drugs. Melanoma-transfected cells were found to be significantly less sensitive to nitrosoureas (carmustine, fotemustine, streptozotocin) and temozolomide with an increase of IC50 values between 3 and 14 when compared to parent cells. No difference in cell survival rates between MGMT-proficient and -deficient cells was observed for melphalan, chlorambucil, busulphan, thiotepa and cisplatin which preferentially induce N7 guanine lesions. Surprisingly, MGMT overexpression increased the sensitivity of CAL77 cells to mitomycin C by approximately 10-fold. Treatment of clonal cell lines with buthionine-[S,R]-sulfoximine (BSO), an inhibitor of γ-glutamylcysteine synthetase which depletes cellular glutathione, completely reversed this unexpected increase in sensitivity to mitomycin C. This observation suggests that glutathione is involved in the sensitivity of MGMT-transfected cells to mitomycin C and may act synergistically with MGMT via an unknown mechanism.

Efficacy of superficial and deep regional hyperthermia combined with systemic chemotherapy and radiotherapy in metastatic melanoma

Response rates of cutaneous-subcutaneous or lymph node metastases of melanoma to systemic chemotherapy are rather low. We report our clinical experience with superficial and deep regional hyperthermia in combination with radiotherapy and/or chemotherapy with carboplatin. We treated 15 patients with metastatic melanoma (6 men, 9 women; age 39-84 years, mean age 60 years) by using superficial or deep regional hyperthermia produced by electromagnetic energy. Superficial hyperthermia was delivered to skin or lymph node metastases in combination with radiochemotherapy in 12 patients, while deep regional hyperthermia was administered with an annular array applicator to lymph node metastases either in combination with radiochemotherapy (1 patient) or with carboplatin alone (2 patients). The clinical response was assessed by clinical evaluation and/or computer tomography and/or ultrasonography at monthly intervals. Both superficial and deep regional hyperthermia was well tolerated. We observed 5 complete local remissions (34%), 6 partial local remissions (40%) and 2 patients with stable disease (13%). The best results were obtained in cutaneous or retroperitoneal metastases. Local response can be achieved in inoperable metastatic melanoma using superficial or deep regional hyperthermia in combination with radiochemotherapy or chemotherapy.

Chemotherapy vs biochemotherapy QOL effect in melanoma

Chemotherapy vs biochemotherapy QOL effect in melanoma

Intumescent cataract after topical mitomycin-C for conjunctival malignant melanoma

Purpose To present the clinical and histologic findings of a patient in whom intumescent cataract developed after successful topical mitomycin-C (MMC) chemotherapy for conjunctival melanoma originating from primary acquired conjunctival melanosis (PAM) with atypia. Design Observational case report; follow-up at 30 months. Methods In a patient with PAM and subsequent conjunctival melanoma that was successfully treated with topical MMC chemotherapy an intumescent cataract developed after two cycles of 0.04% MMC, each for 1 month. Results After MMC chemotherapy pigmentation of the acquired melanosis vanished almost completely. Histopathology of the check-up specimen revealed local tumor control. Six weeks after the completion of the second cycle, an intumescent cataract developed. Cataract surgery was performed uneventfully. The patient was followed up for 30 months. Conclusions In selected cases, topical MMC chemotherapy is effective for treating conjunctival melanoma. Although severe complications are rare and usually transient, development of cataract may be observed. A prospective study should be initiated.

Melanin as a target for melanoma chemotherapy: pro-oxidant effect of oxygen and metals on melanoma viability.

Melanoma cells have a poor ability to mediate oxidative stress, which may be attributed to constitutive abnormalities in their melanosomes. We hypothesize that disorganization of the melanosomes will allow chemical targeting of the melanin within. Chemical studies show that under oxidative conditions, synthetic melanins demonstrate increased metal affinity and a susceptibility to redox cycling with oxygen to form reactive oxygen species. The electron paramagnetic resonance (EPR)-active 5,5'-dimethyl-pyrollidine N-oxide spin adduct was used to show that binding of divalent Zn or Cu to melanin induces a pro-oxidant response under oxygen, generating superoxide and hydroxyl radicals. A similar pro-oxidant behaviour is seen in melanoma cell lines under external peroxide stress. Melanoma cultures grown under 95% O2/5% CO2 atmospheres show markedly reduced viability as compared with normal melanocytes. Cu- and Zn-dithiocarbamate complexes, which induce passive uptake of the metal ions into cells, show significant antimelanoma activity. The antimelanoma effect of metal- and oxygen-induced stress appears additive rather than synergistic; both treatments are shown to be significantly less toxic to melanocytes.

All-trans retinoic acid (atRA) differentially induces apoptosis in matched primary and metastatic melanoma cells -- a speculation on damage effect of atRA via mitochondrial dysfunction and cell cycle redistribution.

All-trans retinoic acid (atRA) has been suggested to exert its cytotoxicity via apoptosis but the mechanisms behind the damage effects have not been fully understood. In this study, we investigated the cytotoxic effects of atRA in eleven primary and matched metastatic cutaneous melanoma cell lines. All the primary and metastatic melanoma cell lines examined expressed the retinoic acid receptors. The cultured melanoma cells treated with atRA showed dysfunction of mitochondria and altered cell cycle distribution, inhibited cell proliferation and apoptosis. The cytotoxic effects of atRA were dose- and time-dependent. The dysfunction of mitochondria and induction of apoptosis were more pronounced in the primary tumor cells than in the metastatic cell lines from the same patients. The data indicate that the cytotoxic effect of atRA was mediated through dysfunction of mitochondria, alterations in cell cycle and induction of apoptosis. Melanoma in early stage may have better response to atRA adjuvant therapy than the melanoma in late stage, suggesting the early utility of atRA in melanoma chemotherapy.

Low-dose immunotherapy does not increase survival when added to chemotherapy for advanced melanoma

Low-dose immunotherapy does not increase survival when added to chemotherapy for advanced melanoma

Protective, anti-tumor monoclonal antibody recognizes a conformational epitope similar to melibiose at the surface of invasive murine melanoma cells.

Polyclonal and monoclonal antibodies (MAbs) have been raised against B16F10 cells collected from growing tumors in vivo or grown in culture media supplemented with normal mouse serum to avoid xenogeneic reactivity. Antibody binding to glutaraldehyde-fixed melanoma cells and Melan A melanocytes was assayed using chemiluminescent-enzyme-linked immunosorbent assay (CL-ELISA) for increased sensitivity. Most of the reactivity of antitumor polyclonal IgG (92%) was inhibitable by a carbohydrate pool consisting of melibiose, mannose, lactose, and sialic acid. Two monoclonal IgG(2a) antibodies, A4 and B11, had their reactivity to melanoma cells completely and specifically inhibited by melibiose. MAb A4 did not bind to alpha-galactosyl residues abundantly expressed in a protozoan mucin used as substrate, and its binding to the tumor cells was not affected by alpha-galactosidase treatment or addition of alpha-methyl-galactopyranoside or raffinose. Recognition of a mimotope similar to melibiose is suggested. MAb is cytotoxic in vitro in a complement-mediated reaction and effectively neutralizes melanoma cells protecting syngeneic mice against tumor development in vivo. This MAb is thus an important tool for further studies on antitumor adjuvant therapy combined with other agents associated with immuno- and chemotherapy of invasive melanoma.

Immunohistochemical analysis of DNA mismatch repair protein and O6-methylguanine-DNA methyltransferase in melanoma metastases in relation to clinical response to DTIC-based chemotherapy

DNA mismatch repair (MMR) deficiency and increased O6-methylguanine-DNA methyltransferase (MGMT) activity have been related to resistance to O6-guanine methylating agents in tumour cell lines. However, the clinical relevance of MMR and MGMT as drug resistance factors is still unclear. In a retrospective study, the expression levels of the MMR proteins, hMSH2, hMSH6 and hMLH1, were analysed by immunohistochemistry in melanoma metastases from 64 patients, who had received dacarbazine (DTIC) based chemotherapy. More than half of the melanoma patients had tumours with no nuclear staining for either hMSH2 or hMSH6 or both, while all tumours showed positive nuclear staining for hMLH1. The response rates were similar in patients with hMSH2 and/or hMSH6 positive tumours to these in patients with negative tumours. By combination of MMR with previously obtained MGMT data, only 2 of 12 responders had tumours with low MGMT and positive MMR expression. Still all except 3 of the non-responders were identified by having either high MGMT expression or absent staining for hMSH2 or hMSH6 or both in their tumours. However, there was no significant correlation of MMR expression alone or combined with MGMT levels with clinical response to DTIC-based chemotherapy in metastatic melanoma.

Cisplatin, Dacarbazine With or Without Subcutaneous Interleukin-2, and Interferon Alfa-2b in Advanced Melanoma Outpatients: Results From an Italian Multicenter Phase III Randomized Clinical Trial

PURPOSE: Phase II and III studies have shown that the addition of interleukin-2 (IL-2) and interferon alfa-2b (IFNα-2b) in multiagent chemotherapy (CT) for advanced melanoma increases overall response (OR), albeit without clear evidence of an improvement in overall survival (OS). Treatment with high-dose IL-2 can cause severe toxicity and is normally administered in an inpatient setting. We conducted a multicenter prospective randomized clinical trial in outpatients with metastatic melanoma to compare CT with biochemotherapy (bioCT) using immunomodulant doses of IL-2 and IFNα-2b. PATIENTS AND METHODS: One hundred seventy-six eligible patients with advanced melanoma were randomized to receive CT (cisplatin and dacarbazine with or without carmustine every 21 days) or bioCT comprising the same CT regimen followed by low-dose subcutaneous IL-2 for 8 days and IFNα2b three times a week, both for six cycles. RESULTS: At a median follow-up of 18 (CT) and 16 (bioCT) months, median OS was 9.5 versus 11.0 months (P = .51), respectively. In the 89 CT-arm patients, 18 ORs (20.2%) (three complete responders [CRs] and 15 partial responders [PRs]) were observed according to World Health Organization criteria. In the 87 bioCT-arm patients, 22 ORs (25.3%) (three CRs and 19 PRs) (P = .70) were recorded. Treatment-related toxicity was fairly similar in both arms. CONCLUSION: The addition of low-dose immunotherapy did not produce a statistically significant advantage in OS, time to progression, or OR. However, the 11-month median OS in the bioCT arm does not differ greatly from the best results with high-dose IL-2–containing regimens reported in the literature. Furthermore, our treatment schedule was carried out on outpatients and had an acceptable level of toxicity.