Dendritic cell generation and CD4+CD25HIGHFOXP3+ regulatory T cells in human head and neck carcinoma during Radio-chemotherapy

Background

Regulatory T cells (Treg) and dendritic cells (DC) play an important role in tumor immunity and immune escape. However, their interplay and the effects of anti-cancer therapy on the human immune system are largely unknown.

Methods

For DC generation, CD14⁺ monocytes were enriched by immunomagnetic selection from peripheral blood of advanced head and neck squamous cell carcinoma (HNSCC) patients and differentiated into immature DC using GM-SCF and IL-4. DC maturation was induced by addition of TNFα. The frequency of CD4⁺CD25^highF0XP3⁺ Treg in HNSCC patients was analyzed before and after radio-chemotherapy (RCT) by four-color flow cytometry.

Results

In HNSCC patients, the frequency of Treg (0.33 ± 0.06%) was significantly (p = 0.001) increased compared to healthy controls (0.11 ± 0.02%), whereas RCT had variable effects on the Treg frequency inducing its increase in some patients and decrease in others. After six days in culture, monocytes of all patients had differentiated into immature DC. However, DC maturation indicated by CD83 up-regulation (70.7 ± 5.5%) was successful only in a subgroup of patients and correlated well with lower frequencies of peripheral blood Treg in those patients.

Conclusion

The frequency of regulatory T cells is elevated in HNSCC patients and may be modulated by RCT. Monocyte-derived DC in HNSCC patients show a maturation deficiency ex vivo. Those preliminary data may have an impact on multimodality clinical trials integrating cellular immune modulation in patients with advanced HNSCC.     

Study of human sperm motility post cryopreservation

Background

Cryopreservation of spermatozoa is a widely used technique to preserve the fertility of males. It can also benefit the armed forces personnel who are to be sent for long recruitments, while leaving their families behind. This study, apart from studying the effects of freezing and thawing, reveals the effect of the post thaw interval on the motility of the human spermatozoa and thus widens the insemination window period.

Methods

A detailed semen analysis was carried out as per the WHO guidelines for 25 samples. The samples were then washed, analysed and frozen in liquid nitrogen. The semen samples were subsequently thawed and similarly analysed after 20 min and 40 min of thawing. This was then followed by statistical analysis of the comparative motilities.

Results

Motility of sperms is found to decrease after cryopreservation. However, the study revealed that after thawing a significant increase in the motility of the sperms was noted with the progression of time (p < 0.05).

Conclusion

By simulating conditions similar to the in vivo conditions for the post thaw semen samples, we can safely wait, confirm the parameters like motility and count, and then inseminate the samples instead of blindly inseminating them immediately after thawing.     

Prevalence of HIV seropositivity among patients with squamous cell carcinoma of the conjunctiva

Objective

To determine the prevalence of HIV seropositivity among patients with squamous cell carcinoma of the conjunctiva.

Methods

All patients with clinical and histopathological confirmation of squamous cell carcinoma seen during a ten year period (July 1999 to June 2009) were tested for HIV (Human Immunodeficiency Virus). The number of patients with squamous cell carcinoma of the conjunctiva who are HIV positive were counted.

Results

A total of thirty-three(33) eyes in thirty-two(32) patients were confirmed histopathologically to have conjunctival squamous cell carcinoma. Their ages ranged from 22 years to 66 years with a mean age of (38.6±11.8) years (SD). The male to female ratio was 1:1.5. Twenty four (75%) of these patients were HIV positive.

Conclusions

Squamous cell carcinoma is associated with the human immunodeficiency virus and is thus a marker for the disease in Benin City, Nigeria.     

Spinning around or stagnation - what do osteoblasts and chondroblasts really like?

Objective

The influcence of cytomechanical forces in cellular migration, proliferation and differentation of mesenchymal stem cells (MSCs) is still poorly understood in detail.

Methods

Human MSCs were isolated and cultivated onto the surface of a 3 × 3 mm porcine collagen I/III carrier. After incubation, cell cultures were transfered to the different cutures systems: regular static tissue flasks (group I), spinner flasks (group II) and rotating wall vessels (group III). Following standard protocols cells were stimulated lineage specific towards the osteogenic and chondrogenic lines. To evaluate the effects of applied cytomechanical forces towards cellular differentiation distinct parameters were measured (morphology, antigen and antigen expression) after a total cultivation period of 21 days in vitro.

Results

Depending on the cultivation technique we found significant differences in both gen and protein expression.

Conclusion

Cytomechanical forces with rotational components strongly influence the osteogenic and chondrogenic differentiation.     

MicroRNA-10b overexpression promotes non-small cell lung cancer cell proliferation and invasion

Background

miRNAs are a class of small non-coding RNA molecules that play an important role in the pathogenesis of human diseases through negative regulation of gene expression. Although miRNA-10b (miR-10b) has been implicated in other tumors, its role in non-small cell lung cancer (NSCLC) is still unknown. The aim of the present study was to investigate the role of miR-10b in NSCLC.

Methods

Expression of miR-10b was analyzed in NSCLC cell line A549 by qRT-PCR. Cell viability was evaluated using Cell Counting Kit (CCK)-8. Cell migration and invasion were evaluated by wound healing assay and transwell assays. Cell cycle and apoptosis analyses were performed. Western blotting was used to predicate the target of miR-10b.

Results

The A549 cell line transfected with the miR-10b exhibited significantly increased proliferation, migration, and invasion capacities when compared with the control cells (P < 0.05). Krüppel-like factor 4 (KLF4) may be indirectly targeted by miR-10b during the proliferation increasing of A549 cells.

Conclusion

In this study, we found that miR-10b is a tumor enhancer in NSCLC. Thus, miR-10b may represent a potential therapeutic target for NSCLC intervention.     

Analysis of chemical composition and bioactive property evaluation of Indian propolis

Objective

To analyze the chemical composition and to evaluate the bioactive potential of hydroalocoholic extract of propolis.

Methods

Ethanol extract of propolis was analyzed by GC-MS, HPTLC and HPLC methods and in vitro antioxidant, anticholinesterase and cytotoxicity assay were performed.

Results

GC-MS analysis revealed the presence of fatty acids, alcohols, and quercetin. Quercetin was identified and quantified by HPTLC and HPLC methods. Dose dependent DPPH and hydroxyl radical scavenging activity of hydroalcoholic extract of propolis was calculated as 16.20 and 34.33 µg/mL respectively. Inhibition of lipid peroxidation was significant and the IC₅₀ value was calculated as 55.56µg/mL. Anticholinesterase activity was less observed. The cytotoxic activity against both breast (MCF-7) and lung cancer (A543) cell lines were significant and the IC₅₀ value was calculated as 10 and 13 µg/mL respectively.

Conclusions

These findings showed that bioactive compounds present in propolis will alleviate many diseases and can be used for better human health.     

Ovarian hemangioma occurring synchronously with contralateral mature cystic teratoma in an 81-year-old patient

Purpose

Ovarian hemangiomas are seen rarely. We present a case of an ovarian hemangioma occurring synchronously with contralateral mature cystic teratoma.

Case history

An 81-year-old woman presented with hypertension and hyponatremia. In ultrasonographic evaluation a pelvic mass was found located at the left ovary. Histologically, a mature cystic teratoma measuring 9.5 × 9 × 8 cm was seen in left ovary. In the right ovary an incidental vascular lesion measuring 3.5 × 1.5 × 1 cm was observed. Final histopathological examination of this lesion demonstrated a hemangioma of cavernous type.

Conclusion

To the best of our knowledge, this is the first ovarian hemangioma case occurring synchronously with contralateral mature cystic teratoma.     

Ferroportin in the progression and prognosis of hepatocellular carcinoma

Background

Hepatocellular carcinoma (HCC) is the fifth most common malignant tumor in men and the seventh in women and understanding the molecular mechanisms of HCC and establishing more effective therapies are critical and urgent issues. Our objective was to study the expression of ferroportin in hepatocellular carcinoma (HCC) tissue samples and the relationship between ferroportin expression and HCC characteristics.

Methods

Sixty HCC tissues and their corresponding para-cancer liver tissues (PCLT) were obtained from sixty HCC patients who had undergone hepatectomy in the Second Xiangya Hospital of Central South University. Ten normal liver tissue samples were also obtained as a control. Immunohistochemistry (IHC) was performed to analyze the ferroportin expression in HCC, and the relationship between ferroportin expression and HCC clinical pathological characteristics also was analyzed. For the evaluation of IHC results, the comprehensive scoring criteria were met according to the staining intensity and the number of positive staining cells. Western blotting was performed to detect the expression level of ferroportin in HCC cell lines.

Results

Ferroportin expression in HCC tissue was significantly lower compared to PCLT and normal liver tissue (P <0.05). Moreover, ferroportin expression was related to liver cancer cell de-differentiation, the severity degree in TNM staging, Edmondson-Steiner grading, intrahepatic metastasis and portal vein invasion. In addition, high expression of ferroportin was observed in normal human liver cell lines L02 and HL7702, whereas weak positive expression and even negative expression of ferroportin were observed in HCC cell lines FOCUS, MHCC-97H, HepG2 and SMMC-7721. Furthermore, among the four kinds of HCC cell lines, the expression level of ferroportin was the lowest in MHCC-97H cells.

Conclusions

Ferroportin expression level declines along with the progression of liver cancer, suggesting that the reduction of ferroportin may serve as an important marker for poor HCC prognosis and as a new therapeutic target.     

Paris saponin VII suppressed the growth of human cervical cancer Hela cells

Background

Saponins of several herbs are known to induce apoptosis in many cancer cells. The present study aimed to investigate the growth inhibitory effect of Paris saponin VII (PS VII), a kind of steroidal saponins from Chonglou (Rhizoma Paridis Chonglou), on the human cervical cancer cell line Hela and the relative molecular mechanisms.

Methods

Hela cells were exposed to different concentrations of PS VII (1 to 100 μM). Inhibition of cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 5-ethynyl-2′-deoxyuridine (EdU) assays. The amount of apoptotic cells was evaluated by flow cytometric analysis. And the protein level of cleaved caspase-3, cleaved caspase-9, Bax, and Bcl-2 was evaluated by Western blot.

Results

The half maximal inhibitory concentration (IC₅₀) value of PS VII for the growth inhibition of Hela cells was 2.62 ± 0.11 μM. PS VII increased the expression of caspase-3, caspase-9, and Bax while decreased that of Bcl-2, suggesting that PS VII may induce apoptosis through intrinsic apoptotic ways.

Conclusions

These data indicate that PS VII has the potential for the treatment of cervical cancer.     

The Anti-cancer Activity of Vernonia divaricata Sw against Leukaemia,Breast and Prostate Cancers In Vitro

Background:

Vernonia divaricata is one of five endemic Vernonia species of Jamaica. The ethnomedicinal uses of other species have been established, however, scientific validation of this species has not yet been done and as such this paper is aimed at identifying the anti-cancer activity of V divaricata against leukaemia, breast and prostate cancer cell lines.

Methods:

Leaves and stems of V divaricata were dried and milled into powder. The crude hexane and methanol extracts of the leaves and stems were obtained and bio-assayed using WST-1 cell proliferation assay against leukaemia, breast and prostate cancer cell lines.

Results:

The crude hexane and methanol extracts of V divaricata were able to significantly retard the growth of the MCF-7 (breast), HL-60 (leukaemia) and the PC-3 (prostate) cancer cell lines. The crude methanol extract of the stem was the strongest, exhibiting anti-proliferation activity with IC₅₀ values of 10.14, 12.63 and 9.894 μg/ml for the HL-60, MCF-7 and PC-3 cancer cell lines, respectively, with the most potent toward prostate cancer.

Conclusion:

The medicinal use of V divaricata as an anti-cancer agent was corroborated as the crude hexane and methanol extracts demonstrated potent anti-proliferation activity and as such hold potential for further research and development into a drug to prevent or treat various cancers.     

Anticancer effect of the extracts from Polyalthia evecta against human hepatoma cell line (HepG2)

Objective

To investigate the anticancer activity of Polyalthia evecta (P. evecta) (Pierre) Finet & Gagnep against human hepatoma cell line (HepG2).

Methods

The anticancer activity was based on (a) the cytotoxicity against human hepatoma cells (HepG2) assessed using a neutral red assay and (b) apoptosis induction determined by evaluation of nuclei morphological changes after DAPI staining. Preliminary phytochemical analysis of the crude extract was assessed by HPLC analysis.

Results

The 50% ethanol-water crude leaf extract of P. evecta (EW-L) showed greater potential anticancer activity with high cytotoxicity [IC₅₀ = (62.8 ± 7.3)µg/mL] and higher selectivity in HepG2 cells than normal Vero cells [selective index (SI) = 7.9]. The SI of EW-L was higher than the positive control, melphalan (SI = 1.6) and the apoptotic cells (46.4 ± 2.6) % induced by EW-L was higher than the melphalan (41.6 ± 2.1)% (P<0.05). The HPLC chromatogram of the EW-L revealed the presence of various kinds of polyphenolics and flavonoids in it.

Conclusions

P. evecta is a potential plant with anticancer activity. The isolation of pure compounds and determination of the bioactivity of individual compounds will be further performed.     

Hypoxia-induced MTA1 promotes MC3T3 osteoblast growth but suppresses MC3T3 osteoblast differentiation

Background

Bone fracture is one of the most common physical injuries in which gene expression and the microenvironment are reprogramed to facilitate the recovery process.

Methods

By specific siRNA transfection and MTT assay, we evaluated the effects of metastasis-associated gene 1 (MTA1) in osteoblast growth. To show the role of MTA1 in osteoblast under hypoxia conditions, by overexpressing and silencing MTA1 expression, we performed mineral deposition and alkaline phosphatase activity assay to observe the differentiation status of osteoblast cells. Real-time PCR and Western blot assays were adopted to detect the expression of certain target genes.

Results

Here, we reported that hypoxia-induced MTA1 expression through hypoxia-induced factor 1 alpha (HIF-1α) and stimulated the growth of osteoblast MC3T3 cells. Silencing of MTA1 through specific siRNA suppressed MC3T3 cell growth and elicited cell differentiation and induced alkaline phosphatase activation and the upregulation of bone morphogenetic protein-2 and osteocalcin.

Conclusions

We found that MTA1 was regulated by HIF-1α in hypoxia circumstance to suppress osteoblast differentiation. These findings provide new insights for bone fracture healing and new strategies to develop potential targets to promote fracture healing.

Electronic supplementary material

The online version of this article (doi:10.1186/s40001-015-0084-x) contains supplementary material, which is available to authorized users.     

Study of 7 Cases of Giant Cell Tumor of Soft Tissue

Background

Primary giant cell tumour of soft tissues is a distinct but uncommon group of neoplasms morphologically identical to osseous giant cell tumor.

Methods

7 patients with painless growing soft tissue mass, having no attachment to underlying bone, were identified in a four years retrospective study from two zonal hospitals of armed forces. Histopathology of these lesions revealed admixture of multinucleated giant cell with mononuclear cells. All patients were treated by surgical resection and followed up for recurrence. Results : There were 5 male and 2 female patients in the age group of 18 to 56 years. All lesions were superficial, circumscribed and involved extremities except one. Histologic transition between benign and malignant lesion was present in only one of the 7 patients that recurred after three months of surgery for which she had to be operated again. 2 of our 7 cases were lost in follow up.

Conclusion

Primary giant cell tumour of soft tissues usually present as a painless mass and needs to be differentiated from other giant cell rich soft tissue tumors. Benign clinical course is expected if the lesion is excised adequately. Its biological behaviour to have low malignant potential is recognized; but this cannot be predicted and metastasis does occur rarely.Key Words: Giant cell tumour of soft tissue, Giant cell tumour of bone, Malignant fibrous histiocytoma     

Primary epididymis malignant triton tumor: case report and review of the literature

Background

Malignant triton tumor (MTT) is a rare and histological complexity characterized by a mixture of peripheral nerve sheath tumors and with rhabdomyoblastic differentiation. It follows a particularly malignant course.

Case presentation

In the present study, we report the first MTT of epididymis. The patient is a 22-year-old male presented with swelling in the left scrotum over a 2-month period. He did not have the history or symptoms of neurofibromatosis type 1. A mass measured about 3 cm × 4 cm was found in the left epididymis by ultrasound and CT scan. It was diagnosed as epididymis tumor and underwent exploration; intraoperative frozen section was diagnosed malignant tumor and treated with radical orchidoepididymectomy. The pathological report was malignant triton tumor. Despite taken high-dose radiation therapy and followed by chemotherapy for four cycles, he was died of progressive disease with multiple metastases 26 months after surgery. The clinic pathologic characteristics and optimal treatment strategy are reviewed.     

Comparative study of automated cryopreservation of red blood cells

Background

Strategic blood reserves are an important component in meeting blood needs and this can be accomplished through the establishment of a frozen blood program.

Methods

One hundred units of packed RBC were glycerolized using the Haemonetics ACP 215 automated cell processor and placed in a −86 °C deep freezer for freezing and storage. Product weight, hematocrit, RBC count, WBC count and hemoglobin were recorded prior to freezing. Twenty five bags were thawed and deglycerolized after every three months starting at one year from the date of first glycerolization In addition to the earlier parameters the bags were assessed for supernatant osmolality, pH, supernatant hemoglobin, ATP levels and supernatant potassium and from these red cell recovery, percentage hemolysis, supernatant glycerol and red cell viability were estimated. All tests were repeated at the end of 7 and 14 days.

Results

The mean red cell recovery was found to be 86.12% on Day 0 and 84% on Day 14. All the bags showed residual glycerol and pH within the acceptable limits upto Day 14. Percentage hemolysis, Mean ATP levels and mean supernatant potassium levels were within acceptable limits upto Day 14. All the units were sterile upto Day 14.

Conclusion

The data in this study showed that the red cells which were glycerolized using the automated platform ACP 215, frozen at −80 °C for more than a year and deglycerolized again using the ACP 215 had excellent viability while being stored at 4 °C during the 14 days of post-thaw storage.     

The therapeutic efficacy of I131-PSCA-mAb in orthotopic mouse models of prostate cancer

Background

Prostate stem cell antigen (PSCA) is upregulated in prostate cancer tissues. Here we aimed to study the therapeutic efficacy of a monoclonal antibody of PSCA-labeled I¹³¹ (I¹³¹-PSCA-mAb) in orthotopic mouse models of prostate cancer.

Methods

The proliferation, apoptosis and invasion abilities of PC-3 and LNCaP cells treated with I¹³¹-PSCA-mAb were measured by methyl thiazolyl tetrazolium assay, flow cytometry and transwell culture, respectively. The human prostate cancer models were established by orthotopic implantation of PC-3 and LNCaP cells in nude mice. I¹³¹-PSCA-mAb distribution and tumor cell apoptosis in the tumor-bearing nude mice were measured.

Results

The inhibitory and apoptosis rates of PC-3 and LNCaP cells treated with I¹³¹-PSCA-mAb reached a maximum of 84%, 80% and 50%, 46%, respectively, which were obviously higher than in the cells treated with I¹³¹-IgG or PSCA-mAb. The invaded number of PC-3 and LNCaP cells treated with I¹³¹-PSCA-mAbe was significantly reduced (P < 0.01) compared with the control group. The ratios of I¹³¹-PSCA-mAb in tumor to intramuscular I¹³¹-PSCA-mAb (T/NT) in tumor-bearing nude mice were increased with time and reached the highest level after 8 h. T/NT stayed above 3.0 after 12 h, and the tumor could still be developed after 24 h. The number of apoptotic cells in tumor tissue of nude mice treated with I¹³¹-PSCA-mAb was larger than that in the control group.

Conclusion

I¹³¹-PSCA-mAb has the potential to become a new targeted therapy drug for the treatment of prostate cancer.     

Research and applications: Comparison and validation of genomic predictors for anticancer drug sensitivity

Background

An enduring challenge in personalized medicine lies in selecting the right drug for each individual patient. While testing of drugs on patients in large trials is the only way to assess their clinical efficacy and toxicity, we dramatically lack resources to test the hundreds of drugs currently under development. Therefore the use of preclinical model systems has been intensively investigated as this approach enables response to hundreds of drugs to be tested in multiple cell lines in parallel.

Methods

Two large-scale pharmacogenomic studies recently screened multiple anticancer drugs on over 1000 cell lines. We propose to combine these datasets to build and robustly validate genomic predictors of drug response. We compared five different approaches for building predictors of increasing complexity. We assessed their performance in cross-validation and in two large validation sets, one containing the same cell lines present in the training set and another dataset composed of cell lines that have never been used during the training phase.

Results

Sixteen drugs were found in common between the datasets. We were able to validate multivariate predictors for three out of the 16 tested drugs, namely irinotecan, PD-0325901, and PLX4720. Moreover, we observed that response to 17-AAG, an inhibitor of Hsp90, could be efficiently predicted by the expression level of a single gene, NQO1.

Conclusion

These results suggest that genomic predictors could be robustly validated for specific drugs. If successfully validated in patients’ tumor cells, and subsequently in clinical trials, they could act as companion tests for the corresponding drugs and play an important role in personalized medicine.     

In vitro chemosensitivity of head and neck cancer cell lines

Background

Systemic treatment of head and neck squamous cell carcinoma (HNSCC) includes a variety of antineoplastic drugs. However, drug-resistance interferes with the effectiveness of chemotherapy. Preclinical testing models are needed in order to develop approaches to overcome chemoresistance.

Methods

Ten human cell lines were obtained from HNSCC, including one with experimentally-induced cisplatin resistance. Inhibition of cell growth by seven chemotherapeutic agents (cisplatin, carboplatin, 5- fluorouracil, methotrexate, bleomycin, vincristin, and paclitaxel) was measured using metabolic MTT-uptake assay and correlated to clinically-achievable plasma concentrations.

Results

All drugs inhibited cell growth in a concentration-dependent manner with an IC50 comparable to that achievable in vivo. However, response curves for methotrexate were unsatisfactory and for paclitaxel, the solubilizer cremophor EL was toxic. Cross-resistance was observed between cisplatin and carboplatin.

Conclusion

Chemosensitivity of HNSCC cell lines can be determined using the MTT-uptake assay. For DNA-interfering cytostatics and vinca alkaloids this is a simple and reproducible procedure. Determined in vitro chemosensitivity serves as a baseline for further experimental approaches aiming to modulate chemoresistance in HNSCC with potential clinical significance.