Treatment Of OSCC Research Articles

Comparing the photodynamic treatment of OSCC between natural and artificial photosensitizers

Purpose: Since Verteporfin is the generally accepted photosensitizer for OSCC and other forms of skin cancer. Its synthetic structure and broad range in absorption has made it harmful to both tumor and healthy cells and tissues. The study aims to see whether a natural and herbal extract like Chlorophyllin, used in primarily bladder cancer, can become an alternative to Verteporfin. Methods: This study uses HSC-2 cell lines and mice models. The cancer cells in vitro and in vivo will be treated with increasing amounts and various durations of Chlorophyllin and Verteporfin. Killing in vitro is measured by MTT assay and in vivo is measured by xenograft tumor size. Positive control is Verteporfin, and negative control is DMSO in vitro or saline solution for the xenograft experiment. Possible results: The three main possible results: (1) Chlorophyllin kills more cancer cells in vitro and in vivo and it has less killed normal tissue. (5) Verteporfin kills more cancer cells but also has more damage to normal tissues when compared to Chlorophyllin. (8) Verteporfin kills more cancer cells in vitro and in vivo and it has less killed normal tissue. Conclusion: The results of the study will determine if natural alternatives are worth investing research into as they can be low-cost extraction, low toxicity, and high selectivity of cells to kill. Chlorophyllin can pave the way to more intensive research of natural photosensitizers in different types of cancers.

Rhopaloic acid A triggers mitochondria damage-induced apoptosis in oral cancer by JNK/BNIP3/Nix-mediated mitophagy

BackgroundOral squamous cell carcinoma (OSCC) is a frequently occurring type of head and neck cancer with a high mortality and morbidity rate. Rhopaloic acid A (RA), a terpenoid derived from sponges, has demonstrated a promising anti-tumor activity, but its effectiveness for treating OSCC remains unknown. PurposeThe aim of this study was to investigate whether RA inhibits the growth of OSCC. MethodsCell viability was evaluated using CCK-8 assays in OSCC cells (Ca9-22, HSC-3 and SAS) and in normal cells (HGF-1) treated with RA. DAPI staining, AO staining, JC-1 staining and immunofluorescence were used to determine apoptosis, mitochondrial membrane potential and autophagy in RA-treated OSCC cells. Protein expression levels were determined by western blotting. Furthermore, the anti-tumor effect of RA was confirmed in vivo using a zebrafish oral cancer xenotransplantation model. ResultsOSCC cells had a significantly reduced viability after RA treatment, but normal cells were not affected. Treatment with RA caused chromatin condensation in OSCC cells, which increased their expression of autophagy- and apoptosis-related proteins. Furthermore, RA caused mitochondrial damage and increased autophagosome formation. Mitophagy was also induced by RA through the JNK/BNIP3/Nix/LC3B pathway. The JNK inhibitor SP600125 prevented both RA-mediated cell death and mitophagy of OSCC cells. A zebrafish xenograft model demonstrated that RA inhibits OSCC growth. ConclusionIn conclusion, RA showed a potent anticancer activity in in vitro and in in vivo oral cancer models by promoting mitochondrial damage-induced apoptosis and mitophagy, which suggests that RA may be useful as a novel and effective treatment for OSCC.

Impact of Interleukin-6 on Oral Squamous Cell Carcinoma Among the South Indian Population.

Introduction Oral squamous cell carcinoma (OSCC) isassociated with high rates of morbidity and mortality. Despite advances in research and treatment, the survival rate of OSCC patients has not changed considerably in recent years. Interleukin-6 (IL-6) is a proinflammatory cytokinethat is involved in the development of various cancers including OSCC. The role of IL-6 is being studied in various cancers;however, its exact mechanism of action in OSCC among the South Indian population has not yet been studied. Thus, the current study aimsto evaluate and assess the impact of IL-6 on OSCC among the South Indian population. Materials and methods Twenty tissues fromOSCC patients and 20 normal tissues surrounding the same area from normal people were gathered from theDepartment of Oral and Maxillofacial Surgery, Saveetha Dental College and Hospital. The tissues were prepared for expression investigations and hematoxylin and eosin staining. The data was presented as mean ± standard deviation, with statistical significance at p<0.05. Results Our results indicate that, in comparison to normal tissues, OSCC samples had increased IL-6 expression levels (p<0.05). Conclusion We conclude that IL-6 has been identified as a key oncogene in the development of tumors and their spread in several types of cancers, including OSCC. Therefore, IL-6 can be used as a potential diagnostic or prognostic biomarker and the use of IL-6 inhibitors can be formulated as a potential treatment forOSCC.

Urolithin A, induces apoptosis and autophagy crosstalk in Oral Squamous Cell Carcinoma via mTOR /AKT/ERK1/2 pathway

BackgroundOral squamous cell carcinoma (OSCC) is the most prevalent malignancy in the world with an alarming rate of mortality. Despite the advancement in treatment strategies and drug developments, the overall survival rate remains poor. Therefore, it is imperative to develop alternative or complimentary anti cancer drugs with minimum off target effects. Urolithin A, a microbial metabolite of ellagic acid and ellagitannins produced endogenously by human gut micro biome is considered to have anti-cancerous activity. However anti tumorigenic effect of urolithin A in OSCC is yet to be elucidated. In this study, we examined whether urolithin A inhibits cell growth and induces both apoptosis and autophagy dependent cell death in OSCC cell lines. PurposeThe present study aims to evaluate the potential of urolithin A to inhibit OSCC and its regulatory effect on OSCC proliferation and invasion in vitro and in vivo mouse models. MethodsWe evaluated whether urolithin A could induce cell death in OSCC in vitro and in vivo mouse models. ResultsFlow cytometric and immunoblot analysis on Urolithin A treated OSCC cell lines revealed that urolithin A markedly induced cell death of OSCC via the induction of endoplasmic reticulum stress and subsequent inhibition of AKT and mTOR signaling as evidenced by decreased levels of phosphorylated mTOR and 4EBP1. This further revealed a possible cross talk between apoptotic and autophagic signaling pathways. In vivo study demonstrated that urolithin A treatment reduced tumor size and showed a decrease in mTOR, ERK1/2 and Akt levels along with a decrease in proliferation marker, Ki67. Taken together, in vitro as well as our in vivo data indicates that urolithin A is a potential anticancer agent and the inhibition of AKT/mTOR/ERK signalling is crucial in Urolithin A induced growth suppression in oral cancer. ConclusionUrolithin A exerts its anti tumorigenic activity through the induction of apoptotic and autophagy pathways in OSCC. Our findings suggest that urolithin A markedly induced cell death of oral squamous cell carcinoma via the induction of endoplasmic reticulum stress and subsequent inhibition of AKT and mTOR signaling as evidenced by decreased levels of phosphorylated mTOR and 4EBP1. Urolithin A remarkably suppressed tumor growth in both in vitro and in vivo mouse models signifying its potential as an anticancer agent in the prevention and treatment of OSCC. Henceforth, our findings provide a new insight into the therapeutic potential of urolithin A in the prevention and treatment of OSCC.

Expression of YT Domain Family Protein in Oral Squamous Cell Carcinoma and Its Correlation with the Prognosis.

Proteins contained in the conserved YTH521-b homologous (YTH) domain, have m6A-dependent RNA binding activity. As an important part of YTH domain family proteins, YTHDF1 and YTHDF3 were shown to be associated with many cancers. This paper aimed to reveal the relationship between the expression of these two proteins and the clinical prognosis of OSCC, providing certain guidance for clinical treatment of OSCC. We detected the expression of YTHDF1 and YTHDF3 in 120 OSCC patients by immunohistochemical analysis. Statistical analysis was used to determine whether the high or low expression of these two genes was significantly associated with age, gender, histological type, clinical stage, or lymph node metastasis. The correlation curve and survival curve of the two genes were produced to evaluate the potential clinical significance. We find the expression of YTHDF1 and YTHDF3 was increased in OSCC tissues compared to adjacent normal tissues. The statistical analysis showed that the expression of YTHDF1 and YTHDF3 was significantly associated with the clinical stage and histological type in OSCC patients. There was also a significant correlation between the expression of YTHDF1 and YTHDF3. A high expression of YTHDF1 and YTHDF3 was related to poor patient prognosis. Our findings suggest that a high expression of YTHDF1 and YTHDF3 may be related to poor patient prognosis.

Abstract 4940: Proteasome and survivin inhibitors synergize to inhibit the growth of oral cancer organoids

Abstract Oral squamous cell carcinoma (OSCC) is the most prevalent subtype of head and neck cancers that has a disproportionately high incidence and mortality in developing countries in South-Central Asia, including Pakistan. The standard treatment for OSCC generally consists of surgery combined with radiotherapy or chemotherapy; however, the overall survival rates have not improved, and only ~50% of patients survive the disease. Therefore, identifying and validating new drug targets can provide new therapeutic options for improved treatment and better prognosis for OSCC patients. The present study aimed to develop an organoid model from patient-isolated primary tumour cells and determine their drug sensitivity profile coupled with mutational analysis. New combinations of targeted drugs with synergistic activity in oral cancer cells were identified through high-throughput screening followed by validation of their activity in oral cancer cells in 2D monolayer and 3D organoid cultures. Results indicate a synergistic activity of proteasome (PR-171) and survivin (YM155) inhibitors in all tested patient-derived tumour cells and four oral cancer cell lines. Moreover, chitosan-based nanocapsules harboring these drugs showed therapeutic efficacy comparable to free drug combinations, indicating a potential for efficient and targeted delivery. Next-generation sequencing data identified missense mutations in eleven cancer-related genes that encode BRAF, EGFR, KRAS, NRAS, HRAS, MEK1, PIK3CA, PTEN, NOTCH1, TP53 and TERT. The ongoing research aims to correlate the drug sensitivity data with the mutational profile of patient-derived tumour cells to find novel targets. In conclusion, we have developed 3D organoids from patient-derived primary tumours and utilized them to validate the synergistic combination between screen-identified proteasome and survivin inhibitors. Our preclinical study provides a rationale for further evaluation of the novel drug combination in vivo and in clinical settings. Citation Format: Muhammad Furqan, Iffat Aleem, Muhammad Tahseen, Raza Hussain, Asif Loya, Muhammad Tariq Mahmood, Philippe Gautier, Kevin Myant, Saira Saleem, Amir Faisal. Proteasome and survivin inhibitors synergize to inhibit the growth of oral cancer organoids. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4940.

The role of acetylation of histone H3 and H4 in oral squamous cell carcinoma

Abstract Oral squamous cell carcinoma (OSCC), which commonly develops in the tongue, buccal mucosa, and epithelium of the oral cavity, is the result of an accumulation of genetic and epigenetic alterations. In OSCC, other epigenetic alterations, such as histone modification were present. Histone modification can predict the prognosis of various cancers. Actually, the regulation and remodeling of chromatin structure caused by histone acetylation was crucial for the growth of tumors. Histone acetylation has received extensive research in OSCC, particularly histone H3 and H4. Additionally, the bulk of OSCC treatment plans focused on controlling the degree of histone acetylation. The acetylation sites of histone H3 and H4 that evolved in OSCC and the effects of these modifications on histone are therefore summarized below. Furthermore, the treatment of OSCC was implemented from the realization of normalizing histone acetylation levels in OSCC, and we summarized target drugs in the resumption of OSCC acetylation level. This review could help with understanding and treating OSCC.

Overexpression of lncRNA HOXA-AS2 promotes the progression of oral squamous cell carcinoma by mediating SNX5 expression

BackgroundOral squamous cell carcinoma (OSCC) is one of the most common head and neck cancers. Long non-coding RNA HOXA-AS2 (lncRNA HOXA-AS2) have been extensively studied in various cancers. However, the expression and function of HOXA-AS2 in OSCC still remain unknown. The aim of this study is to investigate the roles of HOXA-AS2 in OSCC.MethodsOSCC tissues and adjacent normal tissues were obtained from OSCC patients. RT-qPCR and Western blot assays were used to detect the expression of target genes in OSCC tissues or cells. Cells proliferation, migration and invasion were detected by CCK-8 and transwell assays, respectively. The target gene of HOXA-AS2 was confirmed by dual-luciferase reporter gene assay.ResultsWe found that HOXA-AS2 expression was remarkably upregulated in OSCC tissues and cell lines. The downregulation of HOXA-AS2 inhibited cells proliferation, migration and invasion. Our bioinformatics analysis found that HOXA-AS2 can target miR-520c-3p, which was confirmed by dual-luciferase reporter gene assay. The expression of HOXA-AS2 was found to be negatively associated with miR-520c-3p in OSCC tissues. Moreover, sorting nexin 5 (SNX5), a downstream target of miR-520c-3p, was inhibited by miR-520c-3p overexpression. SNX5 was also increased in OSCC tissues and cell lines. Additionally, we found that the higher expression of SNX5 was strongly associated with the tumor grade of OSCC patients in Oncomine database. Most importantly, the knockdown of HOXA-AS2 induced cells apoptosis by promoting autophagy by regulating SNX5.ConclusionHOXA-AS2 served an oncogene and promoted OSCC progression via the miR-520c-3p/SNX5 axis. Thus, HOXA-AS2 may be a new biomarker for diagnosis and treatment of OSCC.

β-Elemene alleviates cisplatin resistance in oral squamous cell carcinoma cell via inhibiting JAK2/STAT3 pathway in vitro and in vivo

ObjectiveTo investigate the effect of β-Elemene (β-Ele) on the cisplatin sensitivity of OSCC cells and its mechanism in vitro and in vivo.MethodsThe human OSCC cell lines Tca-8113 and the cisplatin-resistant cell line Tca-8113-CDDP were cultured with β-Ele or/and cisplatin. The cytotoxicity of cisplatin or β-Ele, cell viability, cell cycles and apoptosis were detected. And the expression of JAK2/STAT3 related protein were detected. The xenograft tumor model of OSCC was established in nude mice and treated with cisplatin and/or β-Ele. The volume and weight of the transplanted tumor was measured, and the expression of p-JAK2 and p-STAT3 and cell apoptosis in the xenograft tumor tissues were detected.ResultsThe combination of β-Ele and cisplatin significantly suppressed the cell proliferation, induced cell cycle arrest, promoted the apoptosis of Tca-8113-CDDP cells, and suppressed the activation of JAK2/STAT3 signaling pathway. The rescue experiments suggested that β-Ele enhanced cisplatin sensitivity via down-regulating JAK2/STAT3 signaling pathway. In vivo, β-Ele and cisplatin synergistically suppressed the tumor growth and induced apoptosis, and down-regulated the expression of p-JAK2 and p-STAT3.Conclusionsβ-Ele inhibits the cell viability and enhances the cisplatin sensitivity of OSCC by blocking the activation of JAK/STAT3 signaling pathway in vitro and in vivo, and the combination of β-Ele and cisplatin maybe a novel treatment for OSCC.

Circ_0014359 promotes oral squamous cell carcinoma progression by sponging miR-149.

This study aimed to explore the role and mechanism of circ_0014359 in the OSCC. We firstly investigated the expression levels of circ_0014359 in OSCC tissues and cell lines. Then, the effects of knocking down circ_0014359 on cellular viability, apoptosis, migration, and invasion of OSCC cell lines were observed by cell counting kit-8 assay, flow cytometry, and transwell assay. Xenografts mouse model was established to explore the in vivo effect of circ_0014359 on the tumor volume and size of OSCC. We found that circ_0014359 was highly expressed in the OSCC tissues and cell lines compared to the normal controls (P<0.05). The expression of circ_0014359 was associated with the survival of patients (P<0.05). For the OSCC cell lines, circ_0014359 knock down induced apoptosis and inhibited migration, invasion, and epithelial-mesenchymal transition of OSCC cells (P<0.001). In vivo, silencing the circ_0014359 blocked the growth of OSCC tumors. The circ_0014359 can directly interact with the micro-RNA-149 (miR-149). Inhibition of miR-149 can rescue the inhibitory effects of circ_0014359 knock down on OSCC cells. The circ_0014359-miR-149 pathway may be a novel target for developing strategies for the diagnosis and treatment of OSCC.

Combined Photothermotherapy and Chemotherapy of Oral Squamous Cell Carcinoma Guided by Multifunctional Nanomaterials Enhanced Photoacoustic Tomography.

BackgroundSquamous cell carcinoma of the head and neck is the sixth most common cancer worldwide, with 40% occurring in the oral cavity. Although the level of early diagnosis and treatment of OSCC has improved significantly, the five-year survival rate of advanced patients remains unsatisfactory. However, the main challenges before us are how to get an early and accurate diagnosis and how to formulate effective treatment. Nanoparticle-based chemo-photothermal therapy has proven to be a promising non-invasive approach to treating oral squamous cell carcinoma treatment.MethodsIn this study, we tried to design and synthesize multifunctional hyaluronic acid (HA) modified gold nanorods/mesoporous silica-based nanoparticles loaded with doxorubicin hydrochloride (DOX) for photoacoustic imaging (PAI) guided cooperative chemo-photothermal therapy.ResultsThe resultant nanocomposite shows favorable biocompatibility, relatively low cytotoxicity, ideal drug loading capability and strong PAI signals. In addition, they showed an excellent photothermal conversion efficiency of 49.02% for photothermal therapy (PTT). Moreover, in vivo and in vitro experiments have shown that synergistic chemo-photothermal therapy has better therapeutic effects than chemotherapy alone or PTT (P < 0.05). After being injected into the CAL-27 tumor-bearing mice, the DOX-AuNRs@mSiO2-HA nanoparticles could accumulate rapidly at the tumor sites and achieve complete ablation of tumors when combined with near-infrared laser irradiation, without obvious side effects on normal tissues.ConclusionOur research provides a solid demonstration of the potential of DOX–AuNRs@mSiO2-HA as a multifunctional platform in PAI-guided photothermal chemotherapy for oral squamous cell carcinoma.

Polydatin, a Glycoside of Resveratrol, Induces Apoptosis and Inhibits Metastasis Oral Squamous Cell Carcinoma Cells In Vitro.

Although various methods, such as surgery and chemotherapy, are applied to the treatment of OSCC, there are problems, such as functional and aesthetic limitations of the mouth and face, drug side effects, and lymph node metastasis. Many researchers are making efforts to develop new therapeutic agents from plant-derived substances to overcome the side effects that occur in oral cancer treatment. Polydatin is known as a natural precursor of resveratrol, and research on its efficacy is being actively conducted recently. Therefore, we investigated whether polydatin can induce apoptosis and whether it affects cell migration and invasion through the regulation of EMT-related factors in OSCC. Polydatin decreased the survival and proliferation rates of CAL27 and Ca9-22 cells, and induced the release of cytochrome c, a factor related to apoptosis, and fragmentation of procaspase-3 and PARP. Another form of cell death, autophagy, was observed in polydatin-treated cells. In addition, polydatin inhibits cell migration and invasion, and it has been shown to occur through increased expression of E-cadherin, an EMT related factor, and decreased expression of N-cadherin and Slug and Snail proteins and genes. These findings suggest that polydatin is a potential oral cancer treatment.

4sc-202 and Ink-128 cooperate to reverse the epithelial to mesenchymal transition in OSCC.

Treatment of oral squamous cell carcinoma remains a challenge due to a high incidence of treatment resistance, which is followed by tumor recrudescence and metastasis to the lymph nodes. Thus, it is important to explore novel inhibitors of OSCC. Here, we aimed to identify drugs that may cooperate with histone deacetylase inhibitors to reverse the EMT, inhibit EMT and cell migration and invasion, and contribute to therapeutic efficacy. We found that treatment with 4sc-202 potently reversed the EMT and thereby inhibited cell migration and invasion in vitro, in part by inducing expression of the FoxO1 tumor-suppressor gene. Furthermore, 4sc-202 also synergized with Ink-128 to inhibit tumor migration and invasion in vitro. Mechanistically, 4sc-202 induced FoxO1 expression, whereas Ink-128 promoted nuclear translocation of FoxO1. Our findings indicated that FoxO1 might reverse the EMT by interacting with Twist1 in OSCC. In conclusion, we identified an effective combination therapy involving class I histone deacetylase and mammalian target of rapamycin complex 1/2 inhibition that effectively blocked the EMT of tumor cells by upregulating FoxO1 expression to inhibit Twist1 transcription. These data have implications for developing new targets for early diagnosis and treatment of OSCC.

Bioinformatics analysis reveals that ANXA1 and SPINK5 are novel tumor suppressor genes in patients with oral squamous cell carcinoma.

BackgroundOral squamous cell carcinoma (OSCC) is a solid tumor of squamous epithelial origin. Currently, surgery is still the main treatment for OSCC, with radiotherapy and chemotherapy as important adjuvant treatments. However, the problem of poor prognosis of OSCC patients still exists in clinical practice. To explore further potential biomarkers or treatment targets in OSCC patients, this study used a high-throughput gene expression database to study the potential molecular mechanisms of OSCC carcinogenesis.MethodsThe GEO database related to OSCC was searched and analyzed using GEO2R. Oncomine and the Human Protein Atlas were used to evaluate the expression level of differentially-expressed genes (DEGs). The cBioPortal dataset was used to analyze the mutations of the potential DEGs and patient survival.ResultsThree GEO datasets, GSE146483, GSE138206, and GSE148944, were downloaded and 7 DEGs were found in common in OSCC tissues. Using Oncomine and the Human Protein Atlas, ANXA1, IL1RN, and SPINK5 were decreased in cancer tissues, while protein levels of APOE and IFI35 were increased accordingly. Interestingly, low levels of ANXA1 and SPINKS were associated with the TNM stage of OSCC patients. No mutations in DEGs were found in OSCC patients, based on the cBioPortal dataset. Survival analysis indicated OSCC patients with high MSR1 had poor overall survival (OS), while low expression of CXCR4, ANXA1, IL1RN, and SPINK5 also predicted poor OS in OSCC patients.ConclusionsOur findings uncovered 7 potential biomarkers of OSCC patients, with ANXA1 and SPINK5 serving as potential tumor suppressor genes in OSCC.

Abstract PO-064: Predicting tumor sensitivity to chemotherapeutic drugs using molecular, cellular and computational methods

Abstract Oral squamous cell carcinoma (OSCC) patients respond poorly to chemotherapy. Predicting anticancer drug response before the treatment of OSCC is a significant challenge. Recently, gene expression profiling and cell-based assays reveals drug sensitivity that could be implemented for drug selection for patients. We analyzed the expression of 11 drug response-related genes in 31 OSCC biopsies collected prior to any treatment, using custom designed PCR array. Further, we investigated drug response pattern of selected anticancer drugs using BH3 apoptotic profiling in the primary cells isolated from OSCC tissues. Then, we correlated the results of drug-response gene expression pattern with apoptotic priming to predict tumor response to chemotherapy. The best performing drug (BPD) and response differences (RD) between the drugs were identified using statistical methods to select the best choice of drug in a personalized manner. Based on the Pierson correlation, we classified OSCC tumors as sensitive, moderately responsive or resistant to chemotherapy. We observed that 2 patients were resistant, 17 patients were moderately responsive and 12 patients were sensitive to chemotherapeutic drugs. We found that up-regulation of genes linked to drug resistance facilitates survival of tumor samples, which was revealed by the percentage of apoptotic priming. Moreover, we found that paclitaxel induced 40-45% apoptotic priming compared to other drugs. Average response difference (RD) analysis showed that 80% of tumors responded well to paclitaxel as compared to other drugs studied that were less effective. The computational prediction of drug responses based on the analysis of multiple clinical features of the tumor will be a novel strategy for accomplishing the long-term goal of precision medicine in oncology. The cancer patients will be benefitted if we computationally account all the tumor characteristics (data) for the selection of most effective and precise therapeutic drug. We developed drug efficacy prediction models using multiple tumor features by employing the statistical methods like multi linear regression (MLR), modified MLR-weighted least square (MLR-WLS) and enhanced MLR-WLS. All the three developed drug efficacy prediction models were then validated using the data of actual OSCC samples (train-test ratio 31: 31) and actual Vs hypothetical samples (train-test ratio 31: 30). The selected best statistical model i.e. enhanced MLR-WLS has then been cross-validated (CV) using 341 theoretical tumor data. Finally, the performances of the models were assessed by the level of learning confidence, significance, accuracy and error terms. We observed that the enhanced MLR-WLS model was the best fit to predict anticancer drug efficacy which may have translational applications. Therefore, drug-response gene expression pattern with the status of apoptotic priming in the tumor cells may be translated to the clinic and which will make impact in precision medicine. Citation Format: N. Rajendra Prasad, Kanimozhi Govindaswamy, G.R. Brindha. Predicting tumor sensitivity to chemotherapeutic drugs using molecular, cellular and computational methods [abstract]. In: Proceedings of the AACR Virtual Special Conference on Artificial Intelligence, Diagnosis, and Imaging; 2021 Jan 13-14. Philadelphia (PA): AACR; Clin Cancer Res 2021;27(5_Suppl):Abstract nr PO-064.

Redefining Adequate Surgical Resection Margins for Oral Squamous Cell Carcinoma: Our Institutional Experience in 5 Consecutive Years

Background: Resection margin status is an important predictor of prognosis in patients with surgically treated OSCC. We introduced the concept of “adequate” versus inadequate” resection margins. Methods: A sample of 87 consecutive patients who underwent surgical treatment for OSCC between 2014 and 2019 were retrospectively examined. Patient demographics, tumour characteristics, adjuvant therapy, recurrence status and patient survival (overall -OS- and disease-free -DFS) were evaluated. According to pathological findings, margins were considered clear (≥5 mm), close (1-5 mm) or involved (<1 mm). Using statistical analysis, a binomial cut-off point was established at 3 mm, and patients were classified into two groups according to primary tumour resection margins: “adequate” (margins ≥3 mm) and “inadequate” (margins <3 mm). Results: Clear surgical margins (≥5 mm) were reported in 72% tumour specimens, close in 12% and involved in 16%. Applying the 3 mm cutt-off, 21% patients were considered to have “inadequate” and 79% “adequate” resections. Adjuvant therapy was provided in 60% of cases, in accordance with Clinical Practice Guidelines. OS rate was 63% and DFS rate 64%. OS was significantly lower (p 0.05). Conclusions: An adequate surgical margin for OSCC could be defined at our institution by ≥3 mm, a close margin (≤2.99 mm) being an adverse risk factor in OSCC survival although further studies are needed to analyse the impact in terms of cancer recurrence rates.

Identification of bioactive cytotoxic compound of red button ginger extracted by solvent fractionation

The antioxidant and antimicrobial activity of Costus woodsonii was previously investigated in our lab. The present study investigated the cytotoxic activity of different plant parts and to identify the bioactive compound(s). MTT assay showed that among different plant parts, rhizome extract exhibited the strongest activity (IC50 of 48 hr treatment: <75.8 µg/ml) against all cell lines tested. Using C18 column chromatography, the isolated compound from rhizome, which demonstrated potent cytotoxic activity (IC50 ≤ 2.4 ± 0.2 µg/ml) against oral squamous cell carcinoma was shown to be dioscin by HPLC, LCMS-MS, and NMR analyses. Our findings also showed that dioscin could be a potential treatment for cisplatin-resistant OSCC and C. woodsonii could serve as a potential commercial source for dioscin. Practical applications Different plant parts of Costus woodsonii have long been used traditionally for medicinal application in water decoction. Nonetheless, the bioactivity study on this plant is scarce. In order to exploit different plant parts of C. woodsonii more scientifically and efficiently for commercial purpose, the present study serves as an important work in understanding the cytotoxic activity of each different parts. This study also provides insight on the effect of extraction solvent on the cytotoxic potential of the selected plant parts. Sequential solvent extraction of rhizome extract followed by fractionation revealed the presence of dioscin, an industrially important steroidal saponin. Overall, this study demonstrated that the rhizome of C. woodsonii may act as a potential source of dioscin.

Abstract A29: Targeted disruption of PI3K/Akt/mTOR signaling pathway induces cell cycle arrest, apoptosis, autophagy, and inhibits inflammation, invasion, and angiogenesis of OSCC cells

Abstract Purpose: The phosphoinositide-3-kinase (PI3K) pathway is frequently altered in human cancer. This has led to the development and study of novel PI3K inhibitors for targeted therapy and also to overcome resistance to radiotherapy. Method: The antitumor effects of PI3K inhibitors (PI-828, PI-103 and PX-866) in terms of cell proliferation, colony formation, induction of apoptosis, cell cycle arrest, invasion, autophagy, and pNF- κB/p65 translocation in SCC-4, SCC-9 and SCC-25 cells were studied by performing MTT, clonogenic, DAPI staining, propidium iodide staining, annexin–V binding, Matrigel invasion, acridine orange staining and immunofluorescence assay. Western blot assay was performed to assess the alteration in the expression of various proteins. Result: PI-828 and PI-103 treatment exhibited dose-dependent inhibition of growth and proliferation of OSCC cells with a concomitant induction of apoptosis, altered cell cycle regulation and decreased invasiveness (p &amp;lt;0.01). PX-866 induced apoptosis, cell cycle arrest, autophagy and a significant decrease in the invasiveness of oral cancer cells as compared to untreated cells (p &amp;lt;0.01). These compounds significantly reduced expression of COX-2, Cyclin-D and VEGF in the treated cells besides cytoplasmic accumulation of pNF-κB/p65 protein. In addition to PI3Kα, inactivation of downstream components, i.e., Akt and mTOR was seen. Conclusion: PI3K inhibitors such as PI-103, PI-828 and PX-866 may be developed as potential chemotherapeutic agents for effective treatment of OSCC and other malignancies associated with activated PI3K/Akt pathway. Note: This abstract was not presented at the conference. Citation Format: Sadhna Aggarwal, Sarah John, Satya N. Das, Suresh C. Sharma. Targeted disruption of PI3K/Akt/mTOR signaling pathway induces cell cycle arrest, apoptosis, autophagy, and inhibits inflammation, invasion, and angiogenesis of OSCC cells [abstract]. In: Proceedings of the AACR Special Conference on Targeting PI3K/mTOR Signaling; 2018 Nov 30-Dec 8; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2020;18(10_Suppl):Abstract nr A29.

Abstract A60: Targeted disruption of PI3K/Akt/mTOR signaling pathway induces cell cycle arrest, apoptosis, and autophagy, and inhibits inflammation, invasion and angiogenesis of OSCC cells

Abstract Purpose: The phosphoinositide-3-kinase (PI3K) pathway is frequently altered in human cancer. This has led to the development and study of novel PI3K inhibitors for targeted therapy and also to overcome resistance to radiotherapy. Method: The antitumor effects of PI3K inhibitors (PI-828, PI-103 and PX-866) in terms of cell proliferation, colony formation, induction of apoptosis, cell cycle arrest, invasion, autophagy, and pNF-κB/p65 translocation in SCC-4, SCC-9 and SCC-25 cells was studied by performing MTT, clonogenic, DAPI staining, propidium iodide staining, annexin-V binding, Matrigel invasion, acridine orange staining and immunofluorescence assay. Western blot assay was performed to assess the alteration in the expression of various proteins. Results: PI-828 and PI-103 treatment exhibited dose-dependent inhibition of growth and proliferation of OSCC cells with a concomitant induction of apoptosis, altered cell cycle regulation and decreased invasiveness (p &amp;lt;0.01). PX-866 induced apoptosis, cell cycle arrest, autophagy and a significant decrease in the invasiveness of oral cancer cells as compared to untreated cells (p &amp;lt;0.01). These compounds significantly reduced expression of COX-2, Cyclin-D and VEGF in the treated cells besides cytoplasmic accumulation of pNF-κB/p65 protein. In addition to PI3Kα, inactivation of downstream components, i.e., Akt and mTOR, was seen. Conclusion: PI3K inhibitors such as PI-103, PI-828 and PX-866 may be developed as potential chemotherapeutic agents for effective treatment of OSCC and other malignancies associated with activated PI3K/Akt pathway. Note: This abstract was not presented at the conference. Citation Format: Sadhna Aggarwal, Sarah John, Suresh C. Sharma, Satya N. Das. Targeted disruption of PI3K/Akt/mTOR signaling pathway induces cell cycle arrest, apoptosis, and autophagy, and inhibits inflammation, invasion and angiogenesis of OSCC cells [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2018 Nov 27-30; Miami Beach, FL. Philadelphia (PA): AACR; Cancer Immunol Res 2020;8(4 Suppl):Abstract nr A60.

Identification and validation of key modules and hub genes associated with the pathological stage of oral squamous cell carcinoma by weighted gene co-expression network analysis.

BackgroundOral squamous cell carcinoma (OSCC) is a major lethal malignant cancer of the head and neck region, yet its molecular mechanisms of tumourigenesis are still unclear.Patients and methodsWe performed weighted gene co-expression network analysis (WGCNA) on RNA-sequencing data with clinical information obtained from The Cancer Genome Atlas (TCGA) database. The relationship between co-expression modules and clinical traits was investigated by Pearson correlation analysis. Furthermore, the prognostic value and expression level of the hub genes of these modules were validated based on data from the TCGA database and other independent datasets from the Gene Expression Omnibus (GEO) database and the Human Protein Atlas database. The significant modules and hub genes were also assessed by functional analysis and gene set enrichment analysis (GSEA).ResultsWe found that the turquoise module was strongly correlated with pathologic T stage and significantly enriched in critical functions and pathways related to tumourigenesis. PPP1R12B, CFD, CRYAB, FAM189A2 and ANGPTL1 were identified and statistically validated as hub genes in the turquoise module and were closely implicated in the prognosis of OSCC. GSEA indicated that five hub genes were significantly involved in many well-known cancer-related biological functions and signaling pathways.ConclusionIn brief, we systematically discovered a co-expressed turquoise module and five hub genes associated with the pathologic T stage for the first time, which provided further insight that WGCNA may reveal the molecular regulatory mechanism involved in the carcinogenesis and progression of OSCC. In addition, the five hub genes may be considered candidate prognostic biomarkers and potential therapeutic targets for the precise early diagnosis, clinical treatment and prognosis of OSCC in the future.