Periodontal disease (PD) is strongly linked to increased risk of oral squamous cell carcinoma (OSCC); however, the specific mechanism through which the development of PD and OSCC is simultaneously promoted remains unclear. This study explored the impact of periodontal pathogens on OSCC progression and the contribution of periodontal pathogen-stimulated OSCC to PD development. The expression of osteoclastogenesis-inducing factors was assessed using quantitative reverse transcription polymerase chain reaction analysis following stimulation of OSCC with lipopolysaccharide (LPS) derived from the periodontal pathogen Porphyromonas gingivalis (Pg), a pathogen commonly responsible for PD. The cell counting kit-8 assay was used to determine the effects of Pg-LPS on OSCC proliferation and drug resistance to cisplatin and 5-fluorouracil. The effects of conditioned medium (CM) derived from Pg-LPS–stimulated OSCC on osteoclastogenesis was evaluated using tartrate-resistant acid phosphatase (TRAP) staining on bone marrow-derived macrophages (BMMs). Pg-LPS administration in SCC-25 and YD-8 OSCC cell lines induced a significant increase in receptor activator of nuclear factor kappa-B ligand mRNA expression; however, it did not affect cell proliferation. Treatment with CM derived from Pg-LPS–stimulated SCC-25 or YD-8 cells markedly enhanced the formation of TRAP-positive multinucleated cells during osteoclast differentiation of BMMs. Altogether, these findings demonstrate that Pg-LPS–stimulated OSCC promoted osteoclastogenesis through a paracrine mechanism.
Probiotic lactic acid bacteria are live microorganisms that provide health benefits when administered in adequate amounts and may exhibit antiproliferative effects on various cancer cell lines, including colon cancer. This study investigates the cytotoxic effects of three Lactobacillus strains - Limosilactobacillus (L.) reuteri VA 102, Ligilactobacillus (L.) animalis VA 105, and Limosilactobacillus (L.) reuteri KCTC 3594 (ATCC 23272) - on mouse colon carcinoma cells (CT-26). Live cells, heat-killed cells, and cell-free supernatant (CFS) of Lactobacillus sp. were prepared and used to treat CT-26 cells at different concentrations. The cytotoxic effect was assessed using the MTT assay. The results indicated that the CFS of all strains significantly reduced the viability of CT-26 cells in a dose-dependent manner, with the VA 102 strain showing the most pronounced effect. Heat-killed cells of L. reuteri VA 102 and L. reuteri KCTC 3594 (ATCC 23272) also reduced cell viability. These findings suggest the potential anticancer properties of these Lactobacillus strains and indicate that CFS and heat-killed cells may offer a safer and more effective alternative to live bacteria for therapeutic applications. Our study contributes to the understanding of the potential of Lactobacillus strains, particularly L. reuteri VA 102, L. reuteri KCTC 3594 (ATCC 23272), and L. animalis VA 105, as possible candidates for cancer treatment and control.
Ophiopogonin D (OPD) is a steroidal glycoside derived from Ophiopogon japonicus , a traditional Chinese medicine with diverse biological activities, including antithrombosis, anti-inflammation, and antitussive effects. To investigate the cellular effects and mechanisms of OPD on oral squamous cell carcinoma, cell viability was explored, and the effects of OPD on cell cycle regulators, apoptotic marker proteins, and key proteins involved in metastasis and signaling pathways were examined by MTT assay and Western blotting in YD38 cells. OPD strongly inhibited cell proliferation and induced caspase-dependent apoptosis of YD38 cells by suppressing the cell cycle and activating caspase-3 and poly ADP ribose polymerase. Additionally, OPD suppressed the expression of vital proteins regulating metastasis and proliferation within the integrin/matrix metalloproteinases/FAK and AKT/PI3K/mTor pathways. Thus, OPD can be a potential treatment candidate for gingival cancer.
Orthokeratinized odontogenic cysts are developmental cysts that occur in the jaw that account for approximately 7%–17% of all cysts in the jaws. Studies have shown that malignant transformation of odontogenic cysts most often occurs in inflammatory cysts, such as periapical cysts, but malignant transformation of orthokeratinized odontogenic cysts has also been reported. In this report, we present an uncommon case of squamous cell carcinoma arising from an orthokeratinized odontogenic cyst.
This study investigated major constituents and anti-inflammatory effects of an ethanol extract of Platycodon grandiflorum leaves. Through HPLC analysis, chlorogenic acid and luteolin-7-O-glucoside were identified as predominant constituents in the ethanol extract. Their anti-inflammatory effects were evaluated using murine macrophage (RAW 264.7 cells) and human lung carcinoma cells (NCI-H292 & A549). The ethanol extract significantly (p<0.01) inhibited the production of nitrite, interleukin-6 (IL-6), and prostaglandin E2 (PGE2) induced by lipopolysaccharide (LPS) in RAW 264.7 cells. Furthermore, the ethanol extract suppressed the expression of cyclooxygenase-2 (COX-2) and inducible NO synthase (iNOS) proteins in RAW 264.7 cells stimulated with LPS. In NCI-H292 and A549 cells, treatment with the ethanol extract significantly (p<0.05) decreased levels of pro-inflammatory cytokines IL-6 and IL-8 induced by IL-1β. The phosphorylation of ERK rather than JNK in the mitogen-activated protein kinase signaling pathway was observed to be a more important mediator in the down-regulation of pro-inflammatory cytokines in NCI-H292 cells. These findings suggest that the ethanol extract of Platycodon grandiflorum leaves containing luteolin-7-O-glucoside exhibits promising anti-inflammatory properties.
Despite advancements in therapeutic approaches, radiotherapy and cisplatin-based chemotherapy remain primary noninvasive treatments for patients with oral squamous cell carcinoma (OSCC). Moreover, the 5-year survival rate for patients with OSCC has remained almost unchanged for several decades, and many side effects of chemotherapy still exist. In this study, three-dimensional (3D) models of OSCC were established using fibroblasts, and the efficacy of various biological inhibitors was evaluated. A culture of epithelial cells with two types of fibroblasts (hTERT-hNOFs and cancer-associated fibroblasts) within a type I collagen matrix resulted in the formation of a continuous layer of tightly packed cells compared to models without fibroblasts. Furthermore, the effects of biological chemicals, including Y27632, latrunculin A, and verteporfin, on these models were investigated. The stratified formation of the epithelial layer and invasion in OSCC 3D-culture models were effectively inhibited by verteporfin, whereas invasion was weakly inhibited by Y27632 and latrunculin. Collectively, the developed OSCC 3D-culture models established with fibroblasts demonstrated the potential for drug screening, with verteporfin showing promising efficacy.
Oral bacterial infections substantially affect the development of various periodontal diseases and oral cancers. However, the molecular mechanisms underlying the association between Fusobacterium nucleatum (F. nucleatum ), a major periodontitis (PT)-associated pathogen, and these diseases require extensive research. Previously, our RNAsequencing analysis identified a few hundred differentially expressed genes in patients with PT and peri-implantitis (PI) than in healthy controls. Thus, in the present study using oral squamous cell carcinoma (OSCC) cells, we aimed to evaluate the effect of F. nucleatum infection on genes that are differentially regulated in patients with PT and PI. Human oral squamous cell carcinoma cell lines OSC-2O, HSC-4, and HN22 were used. These cells were infected with F. nucleatum at a multiplicity of infection of 100 for 3 hours at 37℃ in 5% CO2. Gene expression was then measured using reverse-transcription polymerase chain reaction. Among 18 genes tested, the expression of CSF3, an inflammation-related cytokine, was increased by F. nucleatum infection. Additionally, F. nucleatum infection increased the phosphorylation of AKT, p38 MAPK, and JNK in OSC-20 cells. Treatment with p38 MAPK (SB202190) and JNK (SP600125) inhibitors reduced the enhanced CSF3 expression induced by F. nucleatum infection. Overall, this study demonstrated that F. nucleatum promotes CSF3 expression in OSCC cells through p38 MAPK and JNK signaling pathways, suggesting that p38 MAPK and JNK inhibitors may help treat F. nucleatum-related periodontal diseases by suppressing CSF3 expression.
Mucoepidermoid carcinoma (MEC) is the most prevalent malignant tumor originating from the salivary gland. The gradation of MEC is determined histologically based on cellular composition, with high-grade MEC presenting with distinct characteristics and clinical implications. A 56-year-old male presented with a 3-month history of right facial swelling and a recent onset of pain. A subsequent biopsy confirmed a malignant epithelial tumor, with further imaging assisting in determining the surgical approach. Comprehensive surgery, involving the removal of the right submandibular gland and reconstructive procedures, was undertaken. Histopathological evaluation post-surgery confirmed a high-grade MEC. The differentiation between inflammatory conditions and neoplastic lesions in the orofacial region can be challenging. The gradation of MEC is important in guiding therapeutic decisions. Among various classification systems, the Brandwein system offers detailed histopathological criteria that correlate reliably with clinical features. High-grade MECs, although less frequent, are aggressive and have a lower 5-year survival rate. Accurate histopathological diagnosis is crucial in devising an effective treatment plan. The presented case underlines the importance of a meticulous yet periodic follow-up, considering the aggressive nature of high-grade MECs.
Oral squamous cell carcinoma (OSCC) is the most common type of oral cancer, and it has been steadily increasing in worldwide. Pituitary tumor transforming gene 1 (PTTG1) has been known as oncogene in a verity of cancers. Nevertheless, the expression and role of PTTG1 in OSCC progression remains largely unexplored. In this study, clinical datasets were analyzed to assess the genetic impact of PTTG1 on OSCC progression and to identify its functional roles in OSCC cell lines. We analyzed the expression of PTTG1 in head and neck squamous cell carcinoma (HNSC) and OSCC using databases form the Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO). To investigate the effect of PTTG1 on proliferation and migration abilities in OSCC cell lines, following the knockdown of PTTG1 in HSC-2 and SCC-9 cell lines, we analyzed the proliferation and metastatic abilities of OSCC cells using EdU and Boyden chamber assays. Our database analysis revealed that PTTG1 was significantly overexpressed in tumor tissues compared to normal tissues. Moreover, its expression correlated with clinical parameters of OSCC. In vitro experiments demonstrated that depletion of PTTG1 suppressed the ability of cell proliferation and migration in both HSC-2 and SCC-9 cell lines. In conclusion, our study suggests that PTTG1 may act as an oncogene in OSCC. These findings provide new insights into the mechanisms and clinical implications of PTTG1 expression in OSCC patients.
Patients with oral squamous cell carcinoma (OSCC) generally have an elevated expression of homeobox C6 (HOXC6) gene. We found that HOXC6 was the significantly upregulated gene in hypopharyngeal squamous cell carcinoma (FaDu) cells using RNA-seq analysis. However, it remains unclear whether HOXC6 plays a role in tumor process mechanism. Our study aimed to explore the potential oncogenic role and the detailed molecular mechanism of HOXC6 in FaDu cells. In this study, Sirt1 was validated to be overexpressed in FaDu cells and associated with HOXC6 expression. Overexpression of HOXC6 promoted the cell colony formation, whereas inhibition of Sirt1 by Sirt1 inhibitor EX527 reduced cell proliferation/colony formation and migration, and induced apoptosis in HOXC6 overexpressed FaDu cells. Interestingly, mechanistic study showed that EX527 mediated Sirt1 suppression led to decreased HOXC6 expression and upregulation of Sirt1 significantly increased the expression of HOXC6. HOXC6 was shown to cooperate with Sirt1 to enhance cell survival. We propose that HOXC6 promotes cell growth/colony formation, and that the HOXC6 may be a progression of hypopharyngeal carcinoma by activating Sirt1 pathways.
Sarcomatoid carcinoma is rarely diagnosed as gallbladder cancer. Its aggressive nature, due to the characteristics of both sarcoma and carcinoma, results in a poor prognosis. We report a case of gallbladder sarcomatoid carcinoma in an 82-year-old male who was referred to our hospital for evaluation of gallbladder cancer observed on abdominopelvic computed tomography. The characteristics of the cancer were not confirmed after several imaging modalities. The surgically resected tumor was positive for both cytokeratin and vimentin as revealed via immunohistochemical staining, and a sarcomatoid carcinoma was finally diagnosed. The role of chemotherapy has not yet been identified. Therefore, radiation therapy is planned to reduce the risk of recurrence.
Taxillus yadoriki (Siebold) Dancer is a parasitic plant that grows on camellia trees and is common on Jeju Island. The branches of T. yadoriki have long been used to treat various diseases, including hypertension, diabetes mellitus, viral infections, and arthritis. Although recent studies reported that T. yadoriki has anticancer effects in various human cancer cell lines, including lung cancer, the exact molecular mechanisms supporting its anticancer effects are not well understood. This study aims to assess the anticancer effect of the methanol extract of T. yadoriki branches (METY) on mucoepidermoid carcinoma (MEC) cell lines (MC3 cells and YD15 cells) and explore its mechanism of action. Inhibitory activity of MEC cell proliferation was assessed using the CCK-8 assay. The mechanism of the anticancer effect on METY-treated MC3 cells and YD15 cells was evaluated with Hoechst 33342 stain and Western blot. After treating MC3 cells and YD15 cells with METY for 48 hours, the cytotoxicity of MC3 and YD15 cells increased, and nuclear fragmentation increased in both METY-treated MEC cells. Caspase-3 and cleaved PARP activation demonstrated apoptosis of METY-treated MEC cells. Cell proliferation inhibition with METY was alleviated in METY-treated MEC cells pretreated with zVAD-FMK, supporting the cell proliferation inhibition effect by apoptosis. METY-induced apoptosis in MEC cells occurs through MAP kinase pathways such as p38 and pAkt. MEC cell. METY-induced apoptosis of MEC cells occurs via the p38 and pAkt MAPK pathways. Therefore, METY may be a promising anticancer candidate for the MEC therapeutic strategy.
Oral squamous cell carcinoma (OSCC), which accounts for approximately 90% of oral cancers, has a high rate of local recurrence and a poor prognosis despite improvements in treatment. Exosomes released from OSCC cells promote cell proliferation and metastasis. Although it is clear that the biogenesis of exosomes is mediated by the endosomal sorting complex required for transport (ESCRT) machinery, the gene expression pattern of ESCRT, depending on the cell type, remains elusive. The exosomal release from the human OSCC cell lines, HSC-3 and HSC-4, and their corresponding gefitinib-resistant sub-cell lines, HSC-3/GR and HSC-4/GR, was assessed by western blot and flow cytometry. The levels of ESCRT machinery proteins, including Hrs, Tsg101, and Alix, and whole-cell ubiquitination were evaluated by western blot. We observed that the basal level of exosomal release was higher in HSC-3/GR and HSC-4/GR cells than in HSC-3 and HSC-4 cells, respectively. Long-term gefitinib exposure of each cell line and its corresponding gefitinib-resistant sub-cell line differentially induced the expression of the ESCRT machinery. Furthermore, whole-cell ubiquitination and autophagic flux were shown to be increased in gefitinib-treated HSC-3 and HSC-4 cells. Our data indicate that the expression patterns of the ESCRT machinery genes are differentially regulated by the characteristics of cells, such as intracellular energy metabolism. Therefore, the expression patterns of the ESCRT machinery should be considered as a key factor to improve the treatment strategy for OSCC.
Oral squamous cell carcinoma (OSCC) is the most common type of head and neck cancer and is associated with high recurrence, poor treatment, and low survival rates. Hypoxia-inducible factor-1α (HIF-1α) is a transcription factor that regulates the response to hypoxia, a major factor in the tumor microenvironment that affects tumor development and progression in various cancer types. However, microRNA (miRNA) sequence analysis revealed that only a few miRNAs targeting HIF-1α had been discovered. In the present study, we investigated HIF-1α expression in OSCC and the effect of HIF-1α-targeting miRNAs on the progression and metastatic potential of OSCC. We analyzed public databases to explore which miRNAs target HIF-1α expression. In addition, the expression of proteins involved in the cell cycle, proliferation, and apoptosis in HSC-2 cells was analyzed after miRNA-126 mimic treatment. Furthermore, to investigate the effect of miRNA-126 on the proliferation and invasion ability of OSCC cells, 5-ethynyl-2′-deoxyuridine and Transwell assays were performed. The activities of MMP-2 and MMP-9 were evaluated via gelatin zymography. Our results showed that miRNA-126, which targets HIF-1α, enhances OSCC cell proliferation by regulating the cell cycle and reinforces the cell mobility of OSCC via HIF-1α expression. These findings suggest that miRNA-126 may be a novel marker for OSCC treatment and the development of new tools for patients with OSCC.
Ulva compressa Linnaeus (UCL) is a green algae seaweed that performs photosynthesis and is used as a food material in some Asian regions including Korea. It is known to be the dominant species in copper ion-contaminated seas, and many studies on copper ion resistant mechanisms have been reported. UCL is known to have an excellent antioxidant effect, but limited information is available regarding its other physiological activities. In this study, we investigated the anticancer activity of 30% prethanol extracts of Ulva compressa Linnaeus (30% PeUCL) and the underlying mechanisms of its activity on human FaDu hypopharyngeal squamous carcinoma cells. The 30% PeUCL extracts suppressed FaDu cell viability without affecting normal cells (L929), as determined by MTT and viability assays. Furthermore, the 30% PeUCL extracts induced apoptosis, as determined by DAPI staining. The 30% PeUCL extracts inhibited colony formation effectively as well as wound-healing of FaDu cells, even at noncytotoxic concentrations. In addition, 30% PeUCL extracts induced apoptosis significantly through proteolytic cleavage of caspase-3, -7, and -9, and poly (ADP-ribose) polymerase, and by downregulation of Bcl-2 and upregulation of Bax in FaDu cells, as determined by Western blot analysis. Collectively, these results suggest that the inhibitory effect of 30% PeUCL extracts on the growth of oral cancer cells, colony formation and wound-healing may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, 30% PeUCL extracts can be administered as a natural chemotherapeutic drug for the treatment of human oral cancers.
Background : Mucoepidermoid carcinoma (MEC) is the most common primary epithelial malignant salivary gland tumor in both adults and children. Histological grading of MEC is subjective, but plays an important role in predicting patient prognosis. Immunohistochemistry can accurately diagnose diseases and help with treatment and prognosis. The review of this paper was intended to be helpful in the differential diagnosis of mucinous epidermoid carcinoma. Methods : A PubMed search was carried out. Well-known biomarkers for mucoepidermoid carcinoma were searched in PubMed, and their differences with oral squamous cell carcinoma were compared. Results : When PubMed searched “oral mucoepidermoid carcinoma, biomarker”, a total of 241 papers were found, among which cytokeratin(22), Muc1(membrane-bound mucin1, 9), Muc4( membrane-bound mucin4, 6), Muc5ac (membrane-bound mucin5ac, 4), Muc5b (membrane-bound mucin5b, 3), p63 (15), PCNA (15), p53 (20), EGFR (Epidermal growth factor receptor, 21), c-erbB2 (HER2, 14), and pAKT (2) were searched and investigated. The biomarkers retrieved above were compared with those expressed in squamous cell carcinoma. Conclusion : Due to the above biomarkers, it is possible to classify mucoepidermoid carcinoma and differentiate it from other salivary gland tumors or oral squamous cell carcinoma.
Humulus japonicus (HJ) is a widely used herbal medicine for pulmonary tuberculosis, hypertension, leprosy, and venomous wounds in Asia, particularly in China. Although HJ has certain physiological activities, such as longitudinal bone growth, antioxidation and alleviation of rheumatism, its anticancer activities, other than in colorectal and ovarian cancer, are yet to be studied. In this study, we investigated the anti-cancer activity and mechanism of methanol extracts of HJ (MeHJ) against human FaDu hypopharyngeal squamous carcinoma cells. MeHJ suppressed FaDu cell viability without affecting normal cells (L929), which was demonstrated using the MTT and Live & Dead assays. Furthermore, MeHJ effectively inhibited colony formation of FaDu cells, even at non-cytotoxic concentrations, and significantly induced apoptosis through the proteolytic cleavage of caspase-9, -3, -7, poly (ADP-ribose) polymerase and through the downregulation of BCL-2 and upregulation of BAX in FaDu cells, as determined by DAPI staining, flow cytometry, and western blot analyses. Collectively, these findings suggest that the inhibitory effects of MeHJ on the growth and colony formation of oral cancer cells may be mediated by caspase- and mitochondrial-dependent apoptotic pathways in human FaDu hypopharyngeal squamous carcinoma cells. Therefore, MeHJ has the potential to be used as a natural chemotherapeutic drug against human oral cancer.