Changing cell mechanics—a precondition for malignant transformation of oral squamous carcinoma cells

This work demonstrates that a comprehensive strategy of proteomics identification combined with further validation and detailed functional analysis should be adopted in the field of cancer biomarker discovery. A comparative proteomics approach was utilized to identify differentially expressed proteins in 10 oral squamous carcinoma samples paired with their corresponding normal tissues. A total of 52 significantly and consistently altered proteins were identified with eight of these being reported for the first time in oral squamous carcinoma. Of the eight newly implicated proteins, RACK1 was chosen for detailed analysis. RACK1 was demonstrated to be up-regulated in cancer at both the mRNA and protein levels. Immunohistochemical examination showed that the enhanced expression of RACK1 was correlated with the severity of the epithelial dysplasia as well as clinical stage, lymph node involvement, and recurrence, which are known indicators of a relatively poor prognosis in oral squamous carcinoma patients. RNA interference specifically targeted to silence RACK1 could initiate apoptosis of oral squamous carcinoma cells. Taken together, the results indicate that RACK1 is up-regulated in oral squamous carcinoma, not only being closely related to cell proliferation and apoptosis but also linked to clinical invasiveness and metastasis in carcinogenesis. The observations suggest that RACK1 may be a potential biomarker for early diagnosis, prognosis, and monitoring in the therapy of oral squamous carcinoma. Further this comprehensive strategy could be used for identifying other differentially expressed proteins that have potential to be candidate biomarkers of oral squamous carcinoma.

An imbalance between the activity of matrix metalloproteinases (MMPs) (proteolytic enzymes that degrade protein components of the extracellular matrix) and their inhibitors, the tissue inhibitors of metalloproteinases (TIMPs), may be one of the mechanisms responsible for tumor cell invasion. We have investigated the regulation of MMP-1 and TIMP-1 gene expression in benign and malignant (follicular, anaplastic, and papillary) human thyroid cells. As expected of cells with invasive potential, detectable MMP-1 messenger RNA (mRNA) levels were observed in malignant cells under basal conditions, in contrast to undetectable levels in benign cells. Exposure of these cells, for 1 h, to the active phorbol ester, phorbol 12-myristate 13-acetate (TPA, 100 nmol/L), acting via protein kinase C (PKC), elicited an increase in MMP-1 mRNA, with a peak stimulation after a 3- to 4-h culture period. Epidermal growth factor (EGF, 25 ng/mL), however, acting via protein tyrosine kinase (PTK), stimulated such gene expression in malignant cells but failed to do so in benign cells. TIMP-1 mRNA was not significantly altered by the TPA-PKC, EGF-PTK, or TSH-protein kinase A (PKA) pathways in malignant cells. In benign cells, however, TPA induced a small, though significant, increase in TIMP-1. The MMP-1 stimulation by EGF and lack of TPA-induced rise in TIMP-1 in malignant cells, in sharp contrast to the effects obtained in benign thyrocytes, seems to indicate that the MMP: TIMP balance favors a more extensive extracellular matrix protein breakdown by malignant thyrocytes, as expected of cells exhibiting invasive capacity. TSH (10-500 microU/mL) failed to significantly influence basal MMP-1 or TIMP-1 mRNA levels, but it caused a dose-dependent inhibition in TPA- and EGF-induced MMP-1 mRNA in malignant cells, and TPA-stimulated MMP-1 and TIMP-1 in benign cells. The repressive action of TSH on MMP-1 mRNA was mimicked by forskolin and 8-bromo-cAMP and was abrogated by the PKA inhibitor, H-89, suggesting that the TSH inhibitory action is PKA-mediated. In conclusion, the present study provides novel data on MMP-1 and TIMP-1 gene expression and their modulation by the major signal transduction pathways operating in human thyroid cells. Similar and divergent patterns have emerged in the regulation of such gene expression in benign and malignant human thyrocytes, in many instances in accord with the concept of MMP playing the role of stimulating, and TIMP inhibiting, cell invasion. Although MMP-1 may be just one of the many factors responsible for tumor cell invasion, the present findings demonstrating the possibility, at least in vitro, of repressing MMP gene expression may have important clinical ramifications.

Overexpression of Transforming Growth Factor β1 in Malignant Prostate Cells is Partly Caused by a Runaway of TGF-β1 Auto-induction Mediated Through a Defective Recruitment of Protein Phosphatase 2A by TGF-β Type I Receptor

Nerve Growth Factor and Tyrosine Kinase A Receptor in Oral Squamous Cell Carcinoma: Is There an Association With Perineural Invasion?

This article will review the different modes of action of soluble growth factors in the growth of benign and malignant prostatic cells. Cellular proliferation, growth arrest or even apoptosis requires the participation of appropriate growth factors [1]. Proliferative activities of prostatic epithelial cells, like other cells, are governed by the action of a variety of growth factors. Prostatic growth is traditionally considered to be regulated by androgen, as the prostate is an androgen-sensitive organ, in that the growth and maintenance of the structure and functional integrity are dependent upon the presence of circulating androgen [2,3]. A depletion of this androgenic support, e.g. by bilateral orchidectomy in the host, results in massive apoptosis in prostatic epithelial cells, leading to a rapid rate of tissue involution [4]. Subsequent androgen replacement therapy reactivates prostatic growth. Therefore, androgen is the most potent mitogen to the prostate. It is now apparent that this mitogenic effect of androgen on the prostate is mediated through the action of various growth factors as a consequence of an intricate cell-to-cell interaction, a characteristic feature of benign prostatic growth [5,6]. On the other hand, malignant prostatic growth is characterized by additional mechanisms of cellular proliferation that provide a distinct growth advantage over that of their benign counterparts. Cancer develops as a result of a series of genetic mutations [7]. Prostatic cancer is no exception; however, its progression seems to follow a relatively predictable course, from an androgen-sensitive state to an autonomous state [8]. Sensitivity to androgen in prostatic cancer is mainly due to the presence of androgen receptors in malignant cells, which retain some properties of the benign prostatic cells. However, the manner in which androgen interacts with prostatic cancer cells can be drastically different from that in which it reacts with benign cells. Despite androgen playing an important role in the progression of prostatic cancer, eventually these cancer cells are able to convert from an androgen-responsive growth mode to an androgen-independent mode. Often, a conversion from one state to another is associated with a poorer prognosis and is preceded by the acquisition of new growth advantages within the cancer cells. Again, soluble growth factors are the underlying mechanisms of androgen-sensitive and androgen-insensitive malignant growth. The abnormal growth behaviour in prostatic cancer cells can be manifested by an over-expression of growth-stimulating factor(s) and/or a loss of expression (or function) of growth-suppressive factor(s). The production and action of growth factors in the context of benign and malignant growth of prostatic cells will be the focus of the present discussion.

Cervical lymph nodes with or without metastases from oral squamous carcinoma: a correlation of MRI findings and histopathologic architecture

Does Gas Insufflation during Gynecologic or Urologic Oncologic Laparoscopy Cause Dissemination of Malignant Cells

Backround: Mounting evidence implicates iron in the development of a number of cancers, including breast. The responsible mechanism is poorly understood in breast cancer, although recent work has demonstrated irregular expression of various molecular iron transporters. Materials and Methods: Immunohistochemistry on archived material was used to evaluate differences in expression of iron and haem exporters, importers and storage proteins between breast cancer, DCIS and normal breast tissue. The results were corroborated by Western blotting and RT-qPCR on protein and RNA samples generated from matched normal and cancer specimens collected at mastectomy. The ability of benign and malignant breast cells in vitro to accumulate excess iron in the presence of exogenous iron/haem was evaluated with a ferrozine assay. In addition the effect of exogenous iron and haem on cell phenotype in vitro was assessed in benign and malignant cell lines. Assays utilised included viability (MTT), proliferation (BrdU), migration, anchorage-independent growth (colony formation) and invasive capacity. These assays were then repeated in the presence of iron chelators to investigate their potential utility as a therapeutic modality. Results: Statistically significant increases (p<0.05) were observed in expression of the iron importers DCytB, DMT1 and TfR in breast cancer relative to normal tissue. Levels of the storage protein ferritin were also significantly raised. The iron exporters ferroportin and hephaestin were significantly under-expressed in breast cancer, while hepcidin (which induces exporter degradation) was over-expressed. Levels of the haem importer HCP1 and the exporters BCRP and FLVCR were all significantly down-regulated in cancer relative to normal. Both benign and malignant breast cells were capable of importing excess iron, although only malignant cells could import excess haem. Iron induced increased viability and proliferation in benign cells but had no effect on their migratory or invasive capability, or their ability to form colonies. Haem had no effect on benign cell phenotype. Iron and haem both had significantly positive effects on all these aspects of cell phenotype when introduced to malignant breast cells in vitro. Iron chelators were capable of abrogating these positive effects. Discussion: Archived material and prospectively collected matched tissue pairs demonstrate that breast cancers exhibit altered expression of the proteins involved in transporting iron and haem through the cell. In vitro experiments reveal malignant cells to be capable of taking up both substances, which both appear to drive a more malignant phenotype. These in vitro changes can be reversed through the introduction of iron chelators, implying possible efficacy as a novel therapy in breast cancer. Citation Information: Cancer Res 2011;71(24 Suppl):Abstract nr P5-05-06.

Vascular endothelial growth factor (VEGF) is one of the most potent factors for stimulating angiogenesis, an essential process required for expansion of primary tumour and dissemination of malignant cells. To investigate the possible role of VEGF in facilitating metastasis of prostate cancer via stimulating angiogenesis, we have used Northern and slot blotting, reverse transcription polymerase chain reaction, nucleotide sequence analysis and enzyme-linked immunosorbent assay to compare the VEGF expression in series of human and rat cell lines with either benign or malignant characteristics. We have also employed the chick chorioallantoic membrane (CAM) assay to measure the angiogenic activity of the VEGF derived from both benign and malignant cells. The level of VEGF mRNA expressed in the seven malignant human and rat cell lines is 3.5- to 10-fold higher than that expressed in the benign cell lines. The three metastatic variants, generated by transfection of a benign cell line with DNA extracted from prostate carcinoma cells, expressed 2.5 to 5 times more VEGF mRNA than their parental benign cells. While VEGF 121 and 165 were predominantly expressed by both the benign and malignant cells, the transcript representing VEGF 189 isoform was only detected in the malignant cells. At protein level, three human malignant cell lines produced more VEGF (2.7–7.9 ng ml−1) than the benign cell line (1.3 ng ml−1). CAM assay detected a VEGF-dependent angiogenic activity in the medium from malignant cells, but only a relatively weak VEGF-independent activity in the medium from benign cells. These results demonstrated that malignant cells did over-express VEGF and only the VEGF derived from malignant cells was angiogenically active. Thus, we suggest that the VEGF produced by malignant cells might play an important role in facilitating metastasis of prostatic cancer. © 2000 Cancer Research Campaign

The treatment outcomes for oral sarcomatoid squamous cell carcinoma (OSSCC) are far from satisfactory in our hospital. The aim of this study was to retrospectively summarize the OSSCC cases admitted to our department. From 2003 to 2017, 14 patients were hospitalized and diagnosed with OSSCC. We summarized and analysed the medical histories, diagnostic examinations, treatment strategies, and clinical outcomes of the involved cases. Of the 14 cases, 8 were located in the gingiva. The imageological diagnosis identified the existence of a mass with an infiltrative morphology pre-operatively. The cytopathologic features revealed a malignant neoplasm with a mixture of squamous cell carcinoma (SCC) components and spindle cell neoplastic components. To confirm the diagnosis of OSSCC, the use of the immunohistochemical markers AE1/AE3 and Vimentin were more indicative. Complete follow-up data were available for 12 patients, and at the last follow-up, all 12 of the patients had died. The median overall survival for these patients was 11.67 months (range: 3-24 months). OSSCC patients respond poorly to the strategies solely referring to experiences from oral squamous cell carcinoma (OSCC) treatment. The effective diagnosis and treatment of OSSCC at an early stage is necessary. The treatment for OSSCC still poses a great challenge for clinical oncologists.

Platelet-rich fibrin (PRF), originally used to support soft tissue healing, is also considered a therapeutic option for treating oral lichen planus and leukoplakia. The progression from the two premalignant lesions to the aggressive malignant oral squamous cell carcinoma involves an inflammatory process linked to chemokine expression. Thus, there is a rationale for studying how PRF modulates the expression of chemokines in oral squamous carcinoma cells. To this aim, we expose the oral squamous carcinoma cell line HSC2 to IL1β and TNFα either alone or in the presence of lysates obtained from solid PRF membranes. We report here that in HSC2 cells, PRF lysates significantly reduce the forced transcription of chemokines, e.g., CXCL1, CXCL2, CXCL8, CXCL10, and CCL5. Moreover, PRF lysates attenuate the nuclear translocation of p65 in HSC2 oral epithelial cells when exposed to IL1β and TNFα. PRF lysates further reduce chemokine expression provoked by poly:IC HMW. Even though less pronounced, PRF lysates reduce IL1β- and TNFα-induced chemokine expression in TR146 cells. In primary oral epithelial cells, however, PRF lysates increase the basal expression of CXCL1, CXCL2 and CXCL8. Thus, PRF can exert a biphasic effect on chemokine expression in oral squamous cell carcinoma cell lines and primary oral epithelial cells. These findings suggest that PRF may reduce inflammation in a malignant environment while provoking an immunological response in healthy oral epithelium.

Objective To investigate the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2)protein in oral squamous cell carcinoma tissue,and to explore relationship and vascular endothelial growth factor (VEGF) of them.Methods The expression of iNOS,COX-2 and VEGF in 54 cases of oral mucosa squamous cell carcinoma tissue were detected by immunohistochemistry SP staining.And its correlation to clinicopathologic features were analyzed.Results The positive expression rates of iNOS,COX-2 and VEGF were 66.7％,55.6％,62.5％ respectively.The positive expression of iNOS,COX-2 and VEGF was correlated with pathological grade,TNM stage and tumor recurrence (P ＜ 0.05).There were positive correlations between the expression of iNOS and VEGF,COX-2 and VEGF(P ＜ 0.05).Conclusions The expression of iNOS,COX-2 and VEGF are both higher,they are closely correlated to biological action of oral carcinoma,iNOS,COX-2 participate in VEGF promote the tumor angiogenesis. Key words: Oral squamous cell carcinoma; Carbon monoxide synthase; Cyclooxygenase-2; Vascular endothelial growth factor; Immunohistochemistry

Alkannin (ALK) has anti-inflammatory and anti-tumour activities. We tried to probe the underlying functions of ALK in oral squamous carcinoma (OSCC) cells growth, migration and invasion. OSCC cells viability was investigated after treatment with ALK. Then, BrdU, flow cytometry, Western blot and Transwell assays were executed to appraise proliferation, apoptosis, migration and invasion in OSCC cells with ALK stimulation. The biological functions of miR-9 were explored after miR-9 mimic/inhibitor transfection. The relevance of RECK and miR-9 was predicated by dual luciferase activity assay. JAK1/STAT3 and PI3K/AKT pathways were estimated by Western blot. Tumour formation in vivo was executed by xenograft tumours assay. We found that ALK restrained cell proliferation, facilitated apoptosis, repressed migration and invasion also interdicted JAK1/STAT3 and PI3K/AKT pathways in CAL-27 and SCC-9 cells. miR-9 expression was upgraded in OSCC tissues but decreased in OSCC cells along with ALK administration; meanwhile, overexpressed miR-9 inverted the influences of ALK in OSCC cells growth, migration and invasion. RECK was predicated as a novel target gene of miR-9, and overexpressed RECK hindered JAK1/STAT3 and PI3K/AKT pathways in OSCC cells. ALK prohibited tumour formation in vivo. In conclusion, ALK restrained OSCC cells growth, migration and invasion via adjusting miR-9/RECK axis. Highlights ALK restrains cell growth, migration and invasion in OSCC cells; miR-9 is enhanced in OSCC tissues but is repressed by ALK in OSCC cells; miR-9 inverts the repressive effect of ALK on CAL-27 and SCC-9 cells; RECK is a novel target of miR-9; ALK or RECK hinders JAK1/STAT3 and PI3K/AKT pathways in CAL-27 and SCC-9 cells; ALK prohibits tumour formation in vivo.

Human papillomavirus expression in oral mucosa, premalignant conditions, and squamous cell carcinoma: A retrospective review of the literature

In the present study, box-counting fractal dimensions of benign and malignant cells of breast tumors on cytology material were measured and compared. We selected fine-needle aspiration cytology smears of 14 cases of histopathology-proven infiltrating duct carcinomas and 7 cases of fibroadenoma of the breast. Five cells were randomly selected from each case. Box-counting fractal dimensions of all cells were measured with the help of an image cytometer (Leica, Cambridge, UK), using Quantimet 600 software (Leica). In total, 70 malignant cells and 35 benign cells were studied. The mean fractal dimensions of malignant cells and benign cells were 0.9571 +/- 0.1265 and 0.8354 +/- 0.1367, respectively. There was significant difference in the fractal dimension of malignant and benign cells (P = 0.006, Mann-Whitney nonparametric test). The measurement of fractal dimension may be helpful in discriminating malignant from benign cells. This may be another discriminating feature of malignant cells, along with classic image morphometry based on Euclidean geometry.