FADS1 promotes the progression of laryngeal squamous cell carcinoma through activating AKT/mTOR signaling.

Metabolic abnormality is the major feature of laryngeal squamous cell carcinoma (LSCC), however, the underlying mechanism remain largely elusive. Fatty acid desaturase 1 (FADS1), as the key rate-limiting enzyme of polyunsaturated fatty acids (PUFAs), catalyzes dihomo-gamma-linolenic acid (DGLA) to arachidonic acid (AA). In this study, we reported that the expression of FADS1 was upregulated in LSCC, high FADS1 expression was closely associated with the advanced clinical features and poor prognosis of the recurrent LSCC patients after chemotherapy. Liquid chromatograph-mass spectrometry (LC-MS) analysis revealed that FADS1 overexpression induced greater conversion of DGLA to AA, suggesting an increased activity of FADS1. Similarly, the level of prostaglandin E2 (PGE2), a downstream metabolite of AA, was also elevated in cancerous laryngeal tissues. Functional assays showed that FADS1 knockdown suppressed the proliferation, migration and invasion of LSCC cells, while FADS1 overexpression had the opposite effects. Bioinformatic analysis based on microarray data found that FADS1 could activate AKT/mTOR signaling. This hypothesis was further validated by both in vivo and in vitro assays. Hence, our data has supported the viewpoint that FADS1 is a potential promoter in LSCC progression, and has laid the foundation for further functional research on the PUFA dietary supplementation interventions targeting FADS1/AKT/mTOR pathway for LSCC prevention and treatment.

20(s)-Protopanaxadiol (PPD) increases the radiotherapy sensitivity of laryngeal carcinoma.

Laryngeal carcinoma (LC) is one of the most prevalent malignant tumors in the head and neck area. Due to its high morbidity and mortality, LC poses a serious threat to human life and health. Even with surgical removal, some patients were not sensitive to radiotherapy or experienced transfer or recurrence. 20(s)-Protopanaxadiol (PPD), a natural product from Panax ginseng, has been reported to have cytotoxic effects against several cancer cell lines. However, whether it can improve the radiation sensitivity and the underlying mechanism of PPD's sensitization effect is still unknown. Herein, from in vitro and in vivo experiments, we found that the combination of PPD and radiation not only significantly inhibited proliferation and induced apoptosis, but also suppressed the tumor growth in mouse models. These findings confirmed the role of PPD in enhancing the sensitivity of radiotherapy. Moreover, our work showed that the expression levels of mTOR and its downstream effectors decreased remarkably after PPD addition when compared to radiation only. This result suggested that PPD's excellent synergistic effects with radiation might be associated with the down-regulation of the mTOR signaling pathway in Hep-2 cells.

Retrospective analysis of laser vs other therapeutic modalities for laryngeal papillomatosis: European multicenter study.

PURPOSE: Laryngeal papillomatosis can be an aggressive and potentially life-threatening disease, affecting both children and adults. Local excision is the gold standard of treatment, but recurrences are frequently inevitable. The purpose of this study was to present the experience of three institutions with different therapeutic modalities and discuss them in relation with the relevant literature. METHODS: Sixty patients underwent papilloma resection during the last decade in three institutions (Homburg/Saar and Marburg, Germany and Athens,Greece). Patient data were retrospectively analyzed for therapeutic modalities applied, rate of complications and synechia formation, necessary operations and need for tracheostomy. RESULTS: Carbon dioxide laser therapy was the most common modality applied alone or combined with other treatment modalities. No major complication was observed, while glottic synechia was the most common minor complication in 5 (8.3%) patients. Of the patient cohort 55.6% required reoperation, while no patient required tracheostomy. CONCLUSIONS: Surgical debulking with or without adjuvant treatment remains the mainstay of treatment, which mainly aims to reduce the number and frequency of recurrences since no definitive curative therapy is known so far.

[Apoptosis mechanism of taxol combined with resveratrol on human laryngeal carcinoma Hep-2 cells].

Laryngeal cancer is one of the most common malignant tumors in the respiratory tumors, and its incidence ranks second highest in the respiratory tumors. Resveratrol (Res) is a kind of polyphenols, which can inhibit nucleotides can inhibit the growth of liver cancer cells, gastric cancer cells, pancreatic cells and other tumor cells by inhibiting ribonucleotide reductase in the cells. Taxol (Tax) is a kind of secondary metabolites of Taxus chinensis, which has anti-tumor activity for breast cancer, cervical cancer, ovarian cancer and other tumors by inhibiting cellular microtubule depolymerization. But at present the effects of resveratrol combined with taxol on human laryngeal carcinoma cell strain Hep-2 and their underlying molecular mechanisms are rarely reported. After human laryngeal cancer cell Hep-2 cells were processed with resveratrol (Res) and taxol (Tax), CCK-8 assay was used to evaluate the effect of these two herbs on the proliferation of cancer cells; AO/PI staining and JC-1 were used to detect Hep-1 cells apoptosis; the expression of Bax, Bcl-2, PARP, TRIB3, and XIAP genes was detected by real time quantitative PCR; the activity of caspase-3 and caspase-8 was determined with quantitative fluorescence method. The experimental results showed that compared with Tax, Res medication alone, joint group significantly enhanced inhibition of Hep-2 cells activity, decreased the dosage of Tax, increased the expression of Bax and PARP, TRIB3, reduced the expression of the Bcl-2 and XIAP, and promoted the activity of caspase-3 and caspase-8. The test results showed that compared with the single medication, combined group could significantly increase the inhibitory effect on Hep-2 cells, significantly reduce Tax dosage, increase expressions of Bax, PARP, TRIB3, reduce expressions of Bcl-2, XIAP, and promote activity of caspase-3, caspase-8. This indicated apoptosis of human laryngeal carcinoma cell strain Hep-2 may be induced with Res, Tax, and the combination of these two herbs by mitochondria pathway. It provides valuable clue for further research on combination of Res and Tax for the treatment of laryngeal cancer, and expanding the combined application of Res and Tax.

Phytochemical attributes of four conventionally extracted medicinal plants and cytotoxic evaluation of their extracts on human laryngeal carcinoma (HEp2) cells.

The bioactive composition and cytotoxic and antioxidative/prooxidative effects of four medicinal plants: yarrow (Achillea millefolium L.), hawthorn (Crataegus oxyacantha L.), ground ivy (Glechoma hederacea L.), and olive (Olea europea L.) on human laryngeal carcinoma cell line (HEp2) were investigated. Water extracts of these plants obtained by infusion, maceration, and decoction were characterized for their polyphenol content and antioxidant capacity. Based on the extraction efficiency of polyphenols, the final extracts were obtained whose polyphenolic profile, polysaccharides, mineral content, and cytoprotective activities were determined. The overall highest content of polyphenols and antioxidant capacity was determined in hawthorn, followed by yarrow and ground ivy, and the lowest in olive leaves extract. Phytochemical screening revealed the presence of phenolic acids, as the most abundant bioactive compounds, followed by flavonoids, flavons, and flavonols. All examined medicinal plants reduced the cell viability and reactive oxygen species formation in a dose- and time-dependent manner. Ground ivy and yarrow containing a high content of phenolic acids and polysaccharides were more efficient to decrease the cell survival when compared to olive leaf and hawthorn. Experiments confirmed the importance of polyphenolic composition rather than content of investigated plants and revealed a relationship between the polyphenolic and polysaccharide contents and antioxidant/prooxidant characters of medicinal plants.

Clinical assessment and treatment of the dysfunctional larynx after radiation.

OBJECTIVE: To review the pathophysiology of early and late radiation-related tissue changes, methods to differentiate these changes from disease recurrence, and treatment of these changes in the irradiated larynx. DATA SOURCES: Peer-reviewed publications. REVIEW METHODS: PubMed database search. CONCLUSIONS/IMPLICATIONS FOR PRACTICE: Early and late radiation-related changes in the larynx manifest variably between individual patients. Severe radiation-related tissue changes in the larynx and recurrent malignancy share many clinical characteristics, and the presence of malignancy must be considered in these patients. Positron emission tomography may help select patients who need operative biopsy to rule out recurrence. In patients with a cancer-free but dysfunctional larynx, both surgical and nonsurgical treatment options, including hyperbaric oxygen, are available for attempted salvage. Further investigation is needed before hyperbaric oxygen can be considered standard-of-care treatment for these patients.

Effect of Kushen (Radix Sophorae flavescentis) extract on laryngeal neoplasm Hep2 cells.

OBJECTIVE: To evaluate the effect of fermented extract of Kushen (Radix Sophorae Flavescentis) or non-fermented ESF on laryngeal neoplasms Hep2 cells. METHODS: Use 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay to explore the effect of fermented ESF and non-fermented ESF on Hep2 cells, and detect the mRNA and protein expression level of Bcl-2, Bax and Caspase-3 with reverse transcription polymerase chain reaction (RT-PCR) and Western blot. RESULTS: Both fermented ESF and non-fermented ESF could inhibit laryngeal neoplasm's Hep2 cells, but and the cells did not response to the dilution 1: 320 of fermented ESF, nor to the 1:1280 dilution of non-fermented ESF. As time progressed, the dilution 1:80 of fermented ESF and 1:320 dilution of non-fermented ESF could significantly reduce Bcl-2 mRNA and protein expression and down-regulate Caspase-3 mRNA and protein expression. Bax mRNA and protein were not expressed in Hep2 cells. CONCLUSIONS: Both fermented ESF and non-fermented ESF could inhibit the proliferation of Hep2 cells, and the effect of non-fermented ESF was significantly better than that of the fermented.

Long-term results of RTOG 91-11: a comparison of three nonsurgical treatment strategies to preserve the larynx in patients with locally advanced larynx cancer.

PURPOSE: To report the long-term results of the Intergroup Radiation Therapy Oncology Group 91-11 study evaluating the contribution of chemotherapy added to radiation therapy (RT) for larynx preservation. PATIENTS AND METHODS: Patients with stage III or IV glottic or supraglottic squamous cell cancer were randomly assigned to induction cisplatin/fluorouracil (PF) followed by RT (control arm), concomitant cisplatin/RT, or RT alone. The composite end point of laryngectomy-free survival (LFS) was the primary end point. RESULTS: Five hundred twenty patients were analyzed. Median follow-up for surviving patients is 10.8 years. Both chemotherapy regimens significantly improved LFS compared with RT alone (induction chemotherapy v RT alone: hazard ratio [HR], 0.75; 95% CI, 0.59 to 0.95; P = .02; concomitant chemotherapy v RT alone: HR, 0.78; 95% CI, 0.78 to 0.98; P = .03). Overall survival did not differ significantly, although there was a possibility of worse outcome with concomitant relative to induction chemotherapy (HR, 1.25; 95% CI, 0.98 to 1.61; P = .08). Concomitant cisplatin/RT significantly improved the larynx preservation rate over induction PF followed by RT (HR, 0.58; 95% CI, 0.37 to 0.89; P = .0050) and over RT alone (P < .001), whereas induction PF followed by RT was not better than treatment with RT alone (HR, 1.26; 95% CI, 0.88 to 1.82; P = .35). No difference in late effects was detected, but deaths not attributed to larynx cancer or treatment were higher with concomitant chemotherapy (30.8% v 20.8% with induction chemotherapy and 16.9% with RT alone). CONCLUSION: These 10-year results show that induction PF followed by RT and concomitant cisplatin/RT show similar efficacy for the composite end point of LFS. Locoregional control and larynx preservation were significantly improved with concomitant cisplatin/RT compared with the induction arm or RT alone. New strategies that improve organ preservation and function with less morbidity are needed.

Derris scandens Benth extract potentiates radioresistance of Hep-2 laryngeal cancer cells.

The use of herbal products as radiosensitizers is a promising approach to increase the efficacy of radiotherapy. However, adverse effects related to the use of herbal medicine on radiotherapy are not well characterized. The present study concerns the impact of Derris scandens Benth extract on the radiosensitivity of Hep-2 laryngeal cancer cells. Pretreatment with D. scandens extract prior to gamma irradiation significantly increased clonogenic survival and decreased the proportion of radiation-induced abnormal nuclei of Hep-2 cells. Furthermore, the extract was found to enhance radiation-induced G2/M phase arrest, induce Akt activation, and increase motility of Hep-2 cells. The study thus indicated that D. scandens extract potentiates radioresistance of Hep-2 cells, further demonstrating the importance of cellular background for the adverse effect of D. scandens extract on radiation response in a laryngeal cancer cell line.

Phase 3 randomized trial on larynx preservation comparing sequential vs alternating chemotherapy and radiotherapy.

BACKGROUND: Both induction chemotherapy followed by irradiation and concurrent chemotherapy and radiotherapy have been reported as valuable alternatives to total laryngectomy in patients with advanced larynx or hypopharynx cancer. We report results of the randomized phase 3 trial 24954 from the European Organization for Research and Treatment of Cancer. METHODS: Patients with resectable advanced squamous cell carcinoma of the larynx (tumor stage T3-T4) or hypopharynx (T2-T4), with regional lymph nodes in the neck staged as N0-N2 and with no metastasis, were randomly assigned to treatment in the sequential (or control) or the alternating (or experimental) arm. In the sequential arm, patients with a 50% or more reduction in primary tumor size after two cycles of cisplatin and 5-fluorouracil received another two cycles, followed by radiotherapy (70 Gy total). In the alternating arm, a total of four cycles of cisplatin and 5-fluorouracil (in weeks 1, 4, 7, and 10) were alternated with radiotherapy with 20 Gy during the three 2-week intervals between chemotherapy cycles (60 Gy total). All nonresponders underwent salvage surgery and postoperative radiotherapy. The Kaplan-Meier method was used to obtain time-to-event data. RESULTS: The 450 patients were randomly assigned to treatment (224 to the sequential arm and 226 to the alternating arm). Median follow-up was 6.5 years. Survival with a functional larynx was similar in sequential and alternating arms (hazard ratio of death and/or event = 0.85, 95% confidence interval = 0.68 to 1.06), as were median overall survival (4.4 and 5.1 years, respectively) and median progression-free interval (3.0 and 3.1 years, respectively). Grade 3 or 4 mucositis occurred in 64 (32%) of the 200 patients in the sequential arm who received radiotherapy and in 47 (21%) of the 220 patients in the alternating arm. Late severe edema and/or fibrosis was observed in 32 (16%) patients in the sequential arm and in 25 (11%) in the alternating arm. CONCLUSIONS: Larynx preservation, progression-free interval, and overall survival were similar in both arms, as were acute and late toxic effects.

The role of black seed in the proliferation and biochemical marker levels of Hep-2 cells.

For centuries, people in the Middle East and Southeast Asia have used Nigella sativa, also known as black seed (BS), for its homeopathic effects. The objective of this study was to investigate the role BS might have on the metabolic biomarkers of the Hep-2 cell line. The experimental design entailed six groups of five wells each (50,000 cells). Groups II through VI were treated with BS, lipopolysaccharide (LPS), cortisol, LPS + cortisol, and BS + LPS + cortisol, respectively. Group I was the untreated control group. At the end of 24, 48, and 72 hours, the total cell count, protein and MDA levels were measured by following standard lab protocols. Data collected from this study revealed that Hep-2 cells exposed to LPS and cortisol (group V) resulted in a decrease in cell proliferation compared to the control. BS treatment induced a higher proliferation rate than group V. Similar trends were observed in the metabolic behavior of Hep-2 cells as evidenced by the total protein and MDA levels. The exposure of BS showed a shift in the metabolic pathways. In conclusion, this study showed that exposure to LPS resulted in an alteration in the metabolic function and this phenomenon was further escalated under stressful conditions (increased cortisol exposure). In addition, the use of BS reversed the traumatic condition.

Changes in the noninvasive, in vivo electrical impedance of three xenografts during the necrotic cell-response sequence.

PURPOSE: To investigate the noninvasive, in vivo use of electrical impedance spectroscopy (EIS) as a method for observing the real-time, cellular-level responses of a volume of tissue to therapies. Here, we studied the EIS response during the development and progression of hyperthermia-induced coagulative necrosis in three diverse human xenografts. METHODS AND MATERIALS: A necrotic cell response sequence was selectively induced in three types of subcutaneously-grown human tumor xenografts by applying hyperthermia at 44.5 degrees C. The electrical impedance of the tumors was measured from 100 Hz to 10 MHZ, noninvasively, in vivo during the treatments. From the full spectrum EIS, ratios between resistivities at selected frequencies (p-ratios) were used as indicators of the changes in the electrical impedance spectra of each tumor's cell population. RESULTS: The rho-ratios consistently demonstrated characteristic, early, rapid increases which coincided with cell and organelle swelling typical of early necrosis. These increases subsequently slowed, but no decrease began before the end of treatment, unlike previous, similarly treated, thermo-sensitive EMT6 mouse tumors. This was consistent with the xenograft histology, which revealed ubiquitous, early-stage coagulative necrosis, with no gross plasma membrane damage at the end of treatment. The extent of both the necrosis and p-ratio changes were similar to those seen early in the EMT6 tumor treatment. Within several days after treatment, the xenograft volumes regressed nearly completely, suggesting completion of the cell populations' necrotic response (lysing) during this period. Consistent with this, extended EIS measurements over a 24-h posttreatment period allowed tracking of the necrotic response sequence through this lysing phase for one type of xenograft. CONCLUSION: The change in the electrical impedance of a volume of tumor tissue which occurs during and/or after a hyperthermia treatment can be correlated with the extent of necrosis observed histologically in the cell population.

[Study on relationship between nail-fold microcirculation and histopathology of patients with vocal cord polyp and blood stasis syndrome].

Nail-fold microcirculation of 50 vocal cord polyp patients was observed and compared with hoarseness patients without vocal cord polyp. The results showed that peripheral microcirculation was disturbed in the vocal cord polyp patients. Histopathologic observation of 200 vocal cord polyp patients was made. It was found that principal pathologic change concentrated on the lamina propria mucosae of vocal cord and the most prominent manifestation was disturbance of topical microcirculation of Reinke's layer. In microcirculation aspect, the relationship between polyp of vocal cord and the Blood Stasis Syndrome was confirmed, and pathogenic mechanism of vocal cord polyp was discussed.

[Hungarian-made soya-based enteral nutrition solution in the artificial feeding of patients with pharyngo-laryngeal cancer]. / Hazai elöállítású szójabázisú enterális tápoldat a garat-gégerákos betegek mesterséges táplálásában.

III.-IV. stage, protein-calorie malnourished, pharyngolaryngeal tumorous patients were artificially fed perioperatively. Protein source (soya-protein concentrate, TOFU-soya-protein-cottage cheese) manufactured in Hungary, was administered in enteral feeding through naso-gastric tube. The metabolism, transport and utility of the soya-based diet were investigated by biochemical methods and compared to the previously administered tube-feeding. The method is advised on the base of the favourable experiences of the authors.

[Functional status of the lymphocytes and adrenal cortex in preoperative radiotherapy of patients with laryngeal cancer exposed to different oxygen conditions]. / Funktsional'noe sostoianie limfotsitov i kory nadpochechnikov pri predoperatsionnoi luchevogo terapii bol'nykh rakom gortanii v usloviiakh razlichnogo kislorodnogo rezhima.

The authors presented the results of study of lymphocytic and adrenal cortical function in preoperative radiotherapy of 38 laryngeal cancer patients in different oxygen regimens: in the air and under the conditions of hyperbaric oxygenation (HBO). A method of mean dose fractionation (a single focal dose of 3.3 Gy 3 times a week up to a total dose of 33 Gy) was employed in all the patients. The total lymphocyte count in 1 microliter of the blood, the number of lymphocytes blast transformed as a result of PHA exposure, the number of lymphocytes with a positive reaction to acid phosphatase and blood 11-OCS concentration were examined. Radiation reaction like epitheliitis was shown to be less frequent in patients treated under HBO conditions, the probability of development of epitheliitis in patients with lowered lymphocytic function being higher.

[Function-sparing surgery of laryngeal cancer after radiotherapy]. / O funktsional'no-shchadiashchei khirurgii raka gortani posle luchevoi terapii.

Economical sparing surgery following radiation treatment was carried out in 284 cases of laryngeal cancer. Application of a complex of diagnostic procedures for determining tumor boundaries before and during surgery as well as some improved preoperative radiation treatment (irradiation in a high-pressure oxygen chamber) and surgical methods showed that sparing surgery gives good results, as far as function of the treated organ and patient's survival are concerned.