Decreased GRP78 protein expression is a potential prognostic marker of oral squamous cell carcinoma in Taiwan.

BACKGROUND/PURPOSE: Oral squamous cell carcinoma (OSCC) is an aggressive tumor and its occurrence in Taiwan is closely related to chronic smoking, alcohol consumption, and especially to betel quid chewing. It became the fourth most common malignant tumor of Taiwanese men in 2006. Unfortunately, there are few biomarkers for diagnosis and treatment of this disease. METHODS: To find potential markers, two domestic cell lines (OC2 and OCSL) derived from different grades of OSCC were established and their proteins were compared by global proteomic analysis. The expression differences of GRP78 protein in these two cell lines and clinical samples from OSCC patients were verified. RESULTS: Of the 11 candidate proteins expressed differentially in both cell lines, six [heat shock protein 90 kDa beta member 1 (94 kDa glucose-regulated protein; GRP94), protein disulfide-isomerase precursor, vimentin, tubulin beta-2C chain, 78 kDa glucose-regulated protein precursor (GRP78), and annexin A2] were increased in OC2 cells (low-grade OSCC), and five (heat shock protein 90-beta, annexin A1, stress-induced phosphoprotein 1, elongation factor-2, and integrin alpha-3 precursor) were increased in OCSL cells (high-grade OSCC). Some of these proteins have been previously associated with malignant tumors, but no previous association of GRP78 with OSCC has been reported. GRP78 protein expression in these two OSCC cell lines was confirmed by Western blotting. Immunohistochemical staining of clinical samples from OSCC patients revealed that decreased GRP78 protein expression was significantly correlated with advance tumor stage (p < 0.001) and neck lymph node metastasis (p = 0.001). CONCLUSION: GRP78 protein is a possible biomarker of oral cancer in Taiwan.

The correlation between HIF-1 alpha and VEGF in oral squamous cell carcinomas: Expression patterns and quantitative immunohistochemical analysis.

BACKGROUND: The prognostic predict biomarkers are important in oral squamous cell carcinoma (OSCC). We investigated the expression patterns and quantitation of hypoxia-inducible factor-1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF) in OSCC patients. Among OSCC patients with recurrence and metastasis, the expression percentages of HIF-1α and VEGF also were analyzed. METHODS: Thirty-eight patients (8 hyperkeratosis, 30 oral squamous cell carcinoma) were included in this study. In the follow-up period, 5 OSCC patients had metastasis and 12 OSCC patients had local recurrence. We used computer-assisted image processing to analyze immunohistochemistry (IHC). The quantitative analysis of IHC slides, including upper-layer epithelium (U) and lower-layer epithelium (L), was calculated. RESULTS: We found homogeneous expression of VEGF in the epithelium. However, two patterns of HIF-1α expression were observed: homogeneous and heterogeneous. The highest U + L layer percentage in HIF-1α and VEF expression had significant association in tumor metastasis and recurrence (p<0.001 in HIF-1α and p<0.001 in VEGF). U + L layer HIF-1α expression percentage was >156.4%, and the survival rate was poor (p < 0.001). CONCLUSION: HIF-1α expression was not only influenced by tumor hypoxia, it also reflected tumor cell characteristics. High concentrations of VEGF and HIF-1α may have value as prognostic markers of tumor metastasis and recurrence.

Effect of a nonthermal-atmospheric pressure plasma jet on wound healing: An animal study.

BACKGROUND: The use of a nonthermal plasma (NTP) jet in the treatment of living tissue has been the subject of considerable interest in the field of medical technology, and has the potential to reduce the recovery time of open wounds. We aimed to investigate the wound-healing process by clinical observation, blood tests, and expression of cell adhesion markers and reactive oxygen species in NTP jet-treated rats. METHODS: This study utilized Sprague-Dawley (SD) rats as experimental subjects, and wounds measuring 2 cm × 2 cm were produced on the animals' backs. The experimental group was treated with NTP for 5 min/d for 4 weeks. The NTP was injected in a diffused manner into the cage housing the rats. The SD rats that had not received plasma treatment were designated as the control group. Blood was drawn on Postoperative Day 2, Day 4, and at 3 months. An immunohistochemical stain of E-cadherin and 4-hydroxy-2-nonenal (4-HNE), a reactive oxygen species marker, were evaluated and quantified for analysis using a CMYK color model. RESULTS: A total of 35 SD rats were included in the study (25 in the NTP group and 10 in the control group). Low dose plasma treatment shortened the wound-healing time without damaging organs. In the NTP group, the white blood cell counts at Day 2 post-NTP treatment was not increased significantly more than that in the control group. After quantification of immunohistochemical staining, 4-HNE was increased at Day 14 compared with Day 7 (16.16 ± 12.81% vs. 55.11 ± 8.11%, p < 0.001), and E-cadherin was also increased (52.17 ± 14.96% vs. 70.46 ± 12.78%, p = 0.04) in the NTP group. After comparison of NTP and the control, it was observed that 4-HNE and E-cadherin were increased in the NTP group on Day 14. CONCLUSION: Short-term, low-dose NTP wound treatment was demonstrated to accelerate wound healing in SD rats without vital organ toxicity.

Quantitative physiology and immunohistochemistry of oral lesions.

Angiogenesis and hypoxia are reported to correlate with tumor aggressiveness. In this study, we investigated the potential of optically measured total hemoglobin concentration (THC) and blood oxygen saturation (StO2) as a quantitative measure of angiogenesis and hypoxia in oral lesions with an immunohistochemical comparison. 12 normal subjects and 40 oral patients (22 oral squamous cell carcinoma (SCC), 18 benign/premalignant lesions including 11 verrucous hyperplasia (VH) and 7 hyperkeratosis/parakeratosis (HK)) were studied. The results showed that the THC measurement was consistent with vascular endothelial growth factor (VEGF) and microvessel staining in the stromal area, but StO2 was not associated with HIF-1α. We observed inflammation induced neovascular formation in the stromal area of VH and HK that were likely attributed to higher-than-control THC and StO2 and resulted in no difference in optical measurements between all lesions. However, we found that in majority of SCC, the ratio of THC and StO2 levels between lesions and the surrounding tissues provide potential distinguishing characteristics from VH, which are not visually differentiable from SCC, with a sensitivity, specificity, and accuracy of 91%, 68%, and 76%, respectively.