Isoaaptamine Induces T-47D Cells Apoptosis and Autophagy via Oxidative Stress.

Aaptos is a genus of marine sponge which belongs to Suberitidae and is distributed in tropical and subtropical oceans. Bioactivity-guided fractionation of Aaptos sp. methanolic extract resulted in the isolation of aaptamine, demethyloxyaaptamine, and isoaaptamine. The cytotoxic activity of the isolated compounds was evaluated revealing that isoaaptamine exhibited potent cytotoxic activity against breast cancer T-47D cells. In a concentration-dependent manner, isoaaptamine inhibited the growth of T-47D cells as indicated by short-(MTT) and long-term (colony formation) anti-proliferative assays. The cytotoxic effect of isoaaptamine was mediated through apoptosis as indicated by DNA ladder formation, caspase-7 activation, XIAP inhibition and PARP cleavage. Transmission electron microscopy and flow cytometric analysis using acridine orange dye indicated that isoaaptamine treatment could induce T-47D cells autophagy. Immunoblot assays demonstrated that isoaaptamine treatment significantly activated autophagy marker proteins such as type II LC-3. In addition, isoaaptamine treatment enhanced the activation of DNA damage (γH2AX) and ER stress-related proteins (IRE1 α and BiP). Moreover, the use of isoaaptamine resulted in a significant increase in the generation of reactive oxygen species (ROS) as well as in the disruption of mitochondrial membrane potential (MMP). The pretreatment of T-47D cells with an ROS scavenger, N-acetyl-l-cysteine (NAC), attenuated the apoptosis and MMP disruption induced by isoaaptamine up to 90%, and these effects were mediated by the disruption of nuclear factor erythroid 2-related factor 2 (Nrf 2)/p62 pathway. Taken together, these findings suggested that the cytotoxic effect of isoaaptamine is associated with the induction of apoptosis and autophagy through oxidative stress. Our data indicated that isoaaptamine represents an interesting drug lead in the war against breast cancer.

Antileukemic Scalarane Sesterterpenoids and Meroditerpenoid from Carteriospongia (Phyllospongia) sp., Induce Apoptosis via Dual Inhibitory Effects on Topoisomerase II and Hsp90.

Two new scalarane sesterterpenoids, 12ß-(3'ß-hydroxybutanoyloxy)-20,24-dimethyl-24-oxo-scalara-16-en-25-al (1) and 12ß-(3'ß-hydroxypentanoyloxy)-20,24-dimethyl-24-oxo-scalara-16-en-25-al (2), along with one known tetraprenyltoluquinol-related metabolite (3), were isolated from the sponge Carteriospongia sp. In leukemia Molt 4 cells, 1 at 0.0625 µg/mL (125 nM) triggered mitochondrial membrane potential (MMP) disruption and apoptosis showing more potent effect than 2 and 3. The isolates inhibited topoisomerase IIα expression. The apoptotic-inducing effect of 3 was supported by the in vivo experiment through suppressing the volume of xenograft tumor growth (47.58%) compared with the control. Compound 1 apoptotic mechanism of action in Molt 4 cells was further elucidated through inducing ROS generation, calcium release and ER stress. Using the molecular docking analysis, 1 exhibited more binding affinity to N-terminal ATP-binding pocket of Hsp90 protein than 17-AAG, a standard Hsp90 inhibitor. The expression of Hsp90 client proteins, Akt, p70S6k, NFκB, Raf-1, p-GSK3ß, and XIAP, MDM 2 and Rb2, and CDK4 and Cyclin D3, HIF 1 and HSF1 were suppressed by the use of 1. However, the expression of Hsp70, acetylated tubulin, and activated caspase 3 were induced after 1 treatment. Our results suggested that the proapoptotic effect of the isolates is mediated through the inhibition of Hsp90 and topoisomerase activities.

Induction chemotherapy with docetaxel, cisplatin and fluorouracil followed by surgery and concurrent chemoradiotherapy improves outcome of recurrent advanced head and neck squamous cell carcinoma.

BACKGROUND: Locally recurrent rate of advanced head and neck squamous cell carcinoma (HNSCC) still remains high and the treatment is controversial. PATIENTS AND METHODS: We retrospectively analyzed ninety-three patients with recurrent advanced oral cancer from 2009 to 2013. Sixty-four patients are in the docetaxel with cisplatin and 5'-fluorouracil (TPF) group and the remaining twenty-nine patients are in the cisplatin and 5'-fluorouracil (PF) group. RESULTS: The overall response rate was better in the TPF group (p=0.005) than the PF group. Patients who received induction chemotherapy, TPF, followed by surgery and concurrent chemoradiotherapy (CRT) had better overall survival (OS) (p=0.012) and progression-free survival (PFS) (p=0.038), while patients with prior intra-arterial-infusion-chemotherapy had an adverse impact on OS (p=0.039). CONCLUSION: We showed that induction chemotherapy with TPF, followed by surgery and consolidation CRT, is the ideal choice for recurrent advanced HNSCC with improving response rates and survival. However, prior intra-arterial-infusion-chemotherapy showed an adverse impact.

Interval between intra-arterial infusion chemotherapy and surgery for locally advanced oral squamous cell carcinoma: impacts on effectiveness of chemotherapy and on overall survival.

BACKGROUND: The interval between intra-arterial infusion chemotherapy (IAIC) and surgery was investigated in terms of its effects on survival in patients with locally advanced oral squamous cell carcinoma (OSCC). METHODS: This retrospective study analyzed 126 patients who had completed treatment modalities for stage IV OSCC. All patients were followed up for 3 years. Kaplan-Meier and Cox regression methods were used to determine how survival was affected by general factors, primary tumor volume, TNM stage, and duration of neoadjuvant chemotherapy. RESULTS: In 126 patients treated for locally advanced OSCC by preoperative induction IAIC using methotrexate, multivariate analysis of relevant prognostic factors showed that an IAIC duration longer than 90 days was significantly associated with poor prognosis (hazard ratio, 1.77; P = 0.0259). CONCLUSIONS: Duration of IAIC is a critical factor in the effectiveness of multimodal treatment for locally advanced OSCC. Limiting the induction course to 90 days improves overall survival.

Continuous intra-arterial chemotherapy for downstaging locally advanced oral commissure carcinoma.

BACKGROUND: The purpose of this study was to assess the usefulness, safety, and efficacy of intra-arterial (IA) infusion chemotherapy for patients with locally advanced oral commissure cancer. METHODS: Twenty-one patients with stages III and IV squamous cell carcinoma involving the mouth angle were recruited. Methotrexate (MTX; 50 mg/day) was continuously infused into the external carotid artery for a mean period of 8 days, followed by weekly IA bolus of 25 mg MTX for a mean period of 10 weeks. RESULTS: Thirteen patients (62%) achieved a complete response (CR) and 7 patients (33%) had a partial response (PR). At a median follow-up of 69 months, the estimated 1-year, 3-year, and 5-year survival rates of the patients with CR versus PR were 100% versus 57%, 92% versus 43%, and 80% versus 43%, respectively. CONCLUSION: Our data demonstrate that continuous IA chemotherapy could achieve a competitive acceptable survival rate and improved locoregional control of advanced oral commissure cancer.

Continuous intra-arterial infusion chemotherapy as a palliative treatment for oral squamous cell carcinoma in octogenarian or older patients.

We intend to determine the potential benefits and analyze the outcomes of regional intra-arterial infusion chemotherapy (IAIC) for the octogenarian and older patients with oral cancer. Eighteen patients with oral squamous cell carcinoma were included. They were 12 males and six females with ages ranging from 80 to 96 years. An implantable port-catheter system was used for catheterization. Using a portable pump, methotrexate was given continuously through the external carotid artery for a mean period of 5.5 days (range, 4-7 days) followed by weekly bolus of low dose (25mg) of methotrexate until the clinical condition was stabilized. During the first week of IAIC, all tumors began to reduce in size. Of them 11 (61%) had a complete clinical response and 7 (39%) achieved a partial clinical response, after a mean follow-up period of 26 months (range, 8-72 months). The one- and three-year survival rates were 87% and 56%, respectively. There were no catheter-related complications. The side effects were mild and tolerable. IAIC might be specially indicated for oral cancer elderly with co-morbidity since a radical surgery or extensive radiotherapy can be avoided, and offers an acceptable palliative treatment in such patients.

Advanced penile verrucous carcinoma treated with intra-aortic infusion chemotherapy.

PURPOSE: Traditional treatment for advanced penile verrucous carcinoma is penectomy. This mostly leads to remarkable psychosexual problems and greatly affects quality of life, especially in young patients. To preserve the penis we used intra-aortic infusion chemotherapy for advanced verrucous carcinoma. MATERIALS AND METHODS: From 1991 to 2009 we treated 6 men with penile verrucous carcinoma with continuous intra-aortic infusion of 50 mg methotrexate every 24 hours (average 550 mg, range 400 to 800). Citrovorum factor (6 mg) was given intramuscularly every 6 hours during methotrexate infusion. After continuous methotrexate infusion no further anticancer drug was given to complete responders. Partial responders subsequently received long-term, intermittent, intra-aortic infusion of 50 mg methotrexate or 2 mg mitomycin C plus 250 mg 5-fluorouracil every 1 to 2 weeks until tumors disappeared and all wounds healed. RESULTS: After treatment 4 patients achieved a complete response and were disease-free 3 years 9 months to 17 years 10 months (median 11 years 3 months) after therapy. Two patients had a partial response. The patient with a shaft tumor subsequently underwent total penectomy due to unbearable penile pain 4 years after infusion with various drugs without an appreciable response. He has survived 12 years 5 months after initial treatment. The other patient with glans and prepuce tumors had progression with bilateral inguinal metastases despite 1 1/2 years of infusion therapy. Total penectomy was done. Histological examination of the glans mass revealed moderately differentiated squamous cell carcinoma. Patient condition progressed rapidly and he died 11 months after penectomy. CONCLUSIONS: Intra-aortic infusion chemotherapy is a simple, effective method to treat penile verrucous carcinoma with the uniqueness of preserving the anatomical structure and sexual function in complete responders. For penile verrucous carcinoma, especially in younger patients, intra-aortic infusion chemotherapy may be considered organ sparing treatment before penectomy.

Importance of patient's cooperation in surgical treatment for oral submucous fibrosis.

PURPOSE: The aim of this study was to investigate the results of surgical treatment for oral submucous fibrosis (OSF) in patients who did or did not cooperate with the rehabilitation regimen. PATIENTS AND METHODS: There were 54 patients who had surgical treatment of trismus caused by OSF. Split-thickness skin grafts were used to repair surgical defects after surgery on the fibrous bands. According to postoperative collaboration in the rehabilitation regimen, patients were defined as non-cooperative patients (group I) and cooperative patients (group II). Group I (n = 28) and group II (n = 26) were analyzed separately for changes in preoperative, intraoperative, and postoperative interincisal distances (ID) for at least 6 months after surgery. RESULTS: The mean preoperative ID was 18.9 mm (range, 8 to 25 mm) in group I and 18 mm (range, 7 to 25 mm) in group II. The intraoperative ID increased to an average of 39 mm in group I and 38.5 mm in group II. The mean final follow-up ID was 22 mm in group I and 36.1 mm in group II. When evaluating the changes of ID, only a statistically significant difference was found at final visit between groups. CONCLUSIONS: In our study, we found the patient's cooperation is the primary requirement for success in the treatment of OSF.

Effective eradication of oral verrucous carcinoma with continuous intraarterial infusion chemotherapy.

BACKGROUND: We evaluated the effectiveness of intraarterial methotrexate infusion as a primary therapy for oral verrucous carcinoma (VC). METHODS: Fifteen male patients (mean age, 55 years) were included. By using an implantable port-catheter system and a portable pump, methotrexate was continuously infused to the external carotid artery for a mean period of 7.5 days (50 mg/day), followed by weekly bolus of methotrexate (25 mg) via intraarterial route for a mean period of 10 weeks. RESULTS: The tumor regressed dramatically and disappeared completely after treatment within a mean period of 2.5 months. All patients obtained complete remission and recovered without disfigurement. All patients were alive without disease recurrence at a median follow-up of 43 months. The side effects were tolerable. CONCLUSIONS: The results of our treatment modality for oral VC appear to compete favorably with the results of surgical series and even more with those of radiotherapy series.

The use of artificial dermis for surgical defects in the treatment of oral premalignant lesions.

BACKGROUND: This study investigated the application of an artificial dermis as a substitute for split-thickness skin graft in an oral mucosal defect after excision of a premalignant lesion. METHODS: We examined a total of 125 sites repaired with artificial dermis in 84 patients. Forty-one of 84 patients had two different defects and 43 had a single defect. Of 84 patients, there were 52 oral leukoplakia and 32 oral submucous fibrosis. RESULTS: Most patients chewed betel nuts (97.6%). Most of the premalignant lesions were located in the buccal mucosa (94%). The overall success rate was 100% without any or partial graft loss. The minor wound oozing occurred in three grafts (2.4%). No patient had an immunologic reaction or experienced with significant pain. CONCLUSIONS: Therefore, an artificial dermis may be an alterative to a split-thickness skin graft for patients with oral mucosal defects after removal of premalignant lesions.

Continuous intraarterial infusion chemotherapy for early lip cancer.

Most lip cancers are usually diagnosed and can be treated with good prognosis at an early stage. This study reports our experience of treating seven, previously untreated, patients with lip cancer in stage I or II using intraarterial infusion chemotherapy with a single agent. They were all males with ages ranging from 37 to 69 years. An implantable port-catheter system was used for catheterization. Methotrexate 50mg was infused continuously to the external carotid artery every 24h using a portable pump. Methotrexate was given continuously for a mean period of 7 days (range, 4-10 days) and the total administrated dose of methotrexate for intraarterial infusion ranged from 200 to 500 mg (mean, 350 mg). These seven patients were then given weekly bolus of methotrexate (25mg) via intraarterial route for a range of 6-12 weeks. In every case the tumor regressed dramatically and disappeared completely after treatment within a mean period of 2.5 months. Only one patient died, of non-disease related pneumonia 3 years after infusion therapy. The remaining patients are still alive and no recurrence of carcinoma has been observed at a median follow-up period of 28 months. There was no catheter-related complication. The side effects of infusion chemotherapy were mild and tolerable. Our technique of continuous intraarterial infusion therapy for treatment of early lip cancers seems to be as effective as other standard techniques such as surgery or radiation therapy. This modality achieves good tumor response rates, an excellent cosmetic result, preservation of function and minimal side effects.

Altered p-STAT3 (tyr705) expression is associated with histological grading and intratumour microvessel density in hepatocellular carcinoma.

BACKGROUND: Constitutive activation of signal transducer and activator of transcription 3 at tyrosine residue 705 (p-STAT3 (tyr705)) has been associated with many types of human cancers. However, its potential roles and biological effects in hepatocellular carcinoma (HCC) are not well established. AIM: To explore whether an altered p-STAT3 (tyr705) expression is associated with angiogenesis or proliferation and thereby plays a part in HCC development. METHODS: Paraffin-wax-embedded sections from 69 patients with HCC were collected in this study. Using a semiquantitative immunohistochemical staining method, the expression patterns of p-STAT3 (tyr705) in both HCC lesions and the adjacent non-tumorous liver parenchyma were analysed. The results obtained were further correlated with intratumour microvessel density (MVD), Ki-67 expression, clinicopathological parameters and overall survival. RESULTS: A strong p-STAT3 (tyr705) nuclear staining was observed in 49.3% of HCC lesions, but was reported only in 5.8% of the adjacent non-tumorous liver parenchyma (p<0.001). The expression of p-STAT3 (tyr705) in HCC lesions was significantly and positively correlated with the intratumour MVD (p = 0.002), but not with Ki-67 expression. No significant correlation of p-STAT3 (tyr705) was found in addition to histological grading (p = 0.019). Multivariate Cox regression analysis showed that p-STAT3 (tyr705) expression was a significant predictor of overall survival for HCC (p = 0.036), although the Kaplan-Meier survival curves showed no significant difference between the high and low p-STAT3 (tyr705) expression subgroups. CONCLUSIONS: The results showed that p-STAT3 (tyr705) expression was closely correlated with histological grading and intratumour MVD in HCC. Thus, the potential role of p-STAT3 (tyr705) in HCC development may be through these correlations.

Subungual verrucous carcinoma of the thumb treated by intra-arterial infusion with methotrexate.

BACKGROUND: For preservation of integrity of appearance and function in a 66-year-old male with an extremely rare case of verrucous carcinoma developing from the subungium of his right thumb, intra-arterial infusion with methotrexate was used. OBJECTIVE: To evaluate the effectiveness of arterial infusion with methotrexate in this unusual nail bed cancer. METHODS: Right brachial arterial catheterization and infusion with methotrexate (50 mg) were used every 24 hours together with simultaneous intramuscular injection of 6 mg of leucovorin every 6 hours for 10 days. RESULTS: At 4 years, 8 months after therapy, the patient was in sustained complete remission with a functionally normal right thumb. CONCLUSION: This case study suggests that intra-arterial infusion chemotherapy is a simple and effective method for thumb subungual verrucous carcinoma with the unique advantage of preservation of morphology and functional conditions. It can be considered an effective treatment option.

Comparison of deltopectoralis flap and free radial forearm flap in reconstruction after oral cancer ablation.

Defects in the head and neck resulting from ablation of oral cancer frequently lead to disabling functional and cosmetic deformities. Rehabilitation of patients with oral cancer can be an emotionally and technically challenging endeavor. Selection of the donor tissue is clearly dependent upon the defect that is to be reconstructed. The donor tissue should be of sufficient bulk and pliability to facilitate reestablishing contour and function. Regional flap and free flap have been the mainstays of reconstruction for surgical defects after ablation of oral cancer. The records of 77 patients in whom 49 deltopectoral (DP) flaps and 28 free radial forearm flaps (FRFF) were used were analyzed. The success rate of the DP flap was 89.8% and of the FRFF was 96.4%. The minor complication rate at the transplant site (fistula, dehiscence, hematoma) for the DP flaps was 22.4% and for the FRFFs was 14.3%. Overall complication rate at the transplant site for DP flaps was 32.7% and for the FRFFs was 17.9%. The incidence of partial loss of the donor-site skin graft was 4.1% for the DP flaps and 10.7% for the FRFFs. Overall complication rate at the donor site for DP flaps was 10.2% and for the FRFFs was 39.3%. DP flaps had better aesthetic and functional results at the donor site, than FRFFs. However, the DP flap is a staged procedure and results in prolonged hospitalization. In contrast to FRFF, performing a DP flap avoids the difficult technique of microanastomosis, and therefore reduces the donor-site complication rate and the operation time. Therefore, free flaps can not be routinely substituted for regional flaps, but they represent better reconstructive alternatives for specific and selected patients.

Penile verrucous carcinoma successfully treated by intra-aortic infusion with methotrexate.

OBJECTIVES: Penile verrucous carcinoma is characterized by aggressive local growth and a low metastatic potential. Lower abdominal aortic infusion chemotherapy has the main advantage of delivering a very high concentration of an anticancer drug to the whole pelvic area, including the penis, and is especially suitable to treat penile verrucous carcinoma. METHODS: From 1991 to 2000, 4 cases of penile verrucous carcinoma were treated by continuous intra-aortic infusion with methotrexate (50 mg) every 24 hours, for an average of 10 days. Citrovorum factor (6 mg) was given intramuscularly every 6 hours during the period of methotrexate infusion. RESULTS: After treatment, 3 patients achieved complete remission. They were living disease free 10 years, 10 months, 6 years, 9 months, and 1 year, 8 months after therapy. CONCLUSIONS: Intra-aortic infusion chemotherapy is a simple and effective method for penile verrucous carcinoma with the unique advantage of preserving cosmetic and functional integrity. It may be considered an effective alternative treatment for penile verrucous carcinoma. By using this simple method many unnecessary penectomies can be avoided.