Concentrations Of Cisplatin Research Articles

Pharmacokinetics and Anti-Tumor Efficacy of PEGylated Liposomes Co-Loaded with Cisplatin and Mifepristone.

Although cisplatin is an effective chemotherapy drug used against many types of cancer, it has poor bioavailability, produces severe adverse effects, and frequently leads to tumor resistance. Consequently, more effective formulations are needed. The co-administration of cisplatin with mifepristone, which counters an efflux pump drug-resistance mechanism in tumor cells, has shown important synergism, but without resolving the problem of poor bioavailability. Specificity to tumor tissue and bioavailability have been improved by co-encapsulating cisplatin and mifepristone in a liposomal formulation (L-Cis/MF), which needs further research to complete pre-clinical requirements. The aim of this current contribution was to conduct a pharmacokinetic study of cisplatin and mifepristone in male Wistar rats after administration of L-Cis/MF and the conventional (unencapsulated) formulation. Additionally, the capacity of L-Cis/MF to reduce tumor growth in male nude mice was evaluated following the implantation of xenografts of non-small-cell lung cancer. The better pharmacokinetics (higher plasma concentration) of cisplatin and mifepristone when injected in the liposomal versus the conventional formulation correlated with greater efficacy in controlling tumor growth. Future research on L-Cis/MF will characterize its molecular mechanisms and apply it to other types of cancer affected by the synergism of cisplatin and mifepristone.

Voltammetric sensor for an anti-cancer drug cisplatin based on bismuth nanoparticles/graphene modified glassy carbon electrode

A selective and sensitive voltammetric sensor for the measurement of the chemotherapy drug cisplatin was based on bismuth nanoparticles decorated on a graphene-modified glassy carbon electrode (BiNPs/Gr/GCE). The surface morphologies of electrode modifications were characterized using scanning electron microscopy (SEM). Electrochemical characteristics were evaluated by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Differential pulse voltammetry (DPV) was employed to detect cisplatin oxidation. Under the optimal conditions, oxidation peak currents increased linearly with cisplatin concentration from 6.0 to 180 μM. The detection limit was calculated to be 4.4 μM and the limit of quantification was 14.7 μM. Analytical performance studies demonstrated the good electrode reproducibility and the selectivity of the BiNPs/Gr/GCE towards cisplatin detection. The developed cisplatin sensor was successfully utilized to measure cisplatin in samples of human serum. Recoveries ranged from 89 to 102% and the results from the developed sensor were concordant (P > 0.05) with the results of analysis by high-performance liquid chromatography coupled with ultraviolet detection.

Glycyrrhizic acid enhances the anticancer activity of cisplatin in the human ovarian cancer cell line

This study aimed to investigate the effects of glycyrrhizic acid (GL) on the anticancer activity of cisplatin in A2780 ovarian cancer cells. Cultured A2780 cells were treated with different concentrations of GL and cisplatin individually and in combination. The MTT assay, flow cytometry, wound-healing, and clonogenic assay, were used to determine cell viability, apoptosis, migration, and colony formation, respectively. The effects on superoxide dismutase (SOD) activity were also evaluated. QPCR was used to study the effects of individual and combined treatments with GL and cisplatin on the expression levels of migration genes (MMP2 and MMP9), and some apoptosis pathway genes (caspase-3, −8, −9, and BCL2). A synergistic effect was observed between GL and cisplatin (CI < 1). Combination therapy was significantly more effective in reducing cell viability, suppressing migration and colony formation, inducing apoptosis, and altering gene expression compared to single therapies. GL significantly increased SOD activity. The relative expression of caspase −3, −8, and − 9 increased significantly, and the expression levels of MMP2 and MMP9 decreased significantly in the treated cells. Our results indicate that GL enhances the anticancer activity of cisplatin in the A2780 cell line. Therefore, the combination of GL and cisplatin can be proposed as a promising therapeutic strategy for ovarian cancer.

Role of MLH1 and MSH2 deficiency in the development of tumorigenesis and chemo-tolerance of cervical Carcinoma: Clinical implications

Cervical cancer (CACX) is one of the top causes of cancer death in women globally. The involvement of several cellular pathways in carcinogenesis is still poorly understood. Here, we focused to evaluate the contributory role of Mismatch Repair (MMR) pathway genes-MLH1 and MSH2 in CACX and their association with chemo-tolerance of the disease. For this purpose, molecular profiles (expression/promoter methylation/deletion) of the genes were analysed in both normal cervical epithelium and tumour tissue, also validated in in-silico dataset as well. Later on, prognostic importance of the genes was identified through analysis of their methylation/expression status in plasma DNA of circulating tumour cells (CTCs) and cisplatin-tolerant CACX cell lines respectively. It was found that the expression profile of MLH1 and MSH2 genes was considerably reduced from undifferentiated basal-parabasal layers of normal cervical epithelium towards progression of the disease. Further analysis showed that frequent deletion [34–48%] and promoter methylation events [28–46%] of the genes were the plausible reasons for their reduced expression during tumorigenesis. Incidentally, the prevalence of MLH1 [32%] and MSH2 [27%] promoter methylation found in CTCs of plasma of the clinically advanced CACX patients implicated their prognostic importance of the disease. In addition, the patients having high alterations of those genes resulted in poor patient outcomes even after the therapy. In in-depth analysis of this result in cisplatin-tolerant CACX cell lines, we discovered that increased promoter methylation frequency of those genes at higher concentrations of cisplatin and gradual accumulation of the cells in the G2/M phase of the cell cycle were the rational causes for their reduced expression and MMR deficiency in the system. Hence, it is possible to conclude that the gradual down-regulation of MLH1 and MSH2 proteins may be a key event for MMR pathway inactivation in CACX. This might also be associated with chemo-tolerance and overall poor survival among the patients.

Triple-layered scaffold containing cisplatin and curcumin applied for cancerous bone regeneration.

Novel drug delivery system was prepared and evaluated as for application in cancerous bone treatment. It had three stacked layers, including a drug-free layer, a cisplatin-loaded layer, and a curcumin-containing layer. Our previously characterized biomaterials, namely Zr-hydroxyapatite (Zr-HA) and alkaline-treated polycaprolactone (modified-PCL), were used as major components of the drug carrier. Polar-polar interactions between cisplatin and Zr-HA were modulated by the modified-PCL included, leading to increase of cisplatin release. Using β-cyclodextrin (β-CD) to entrap curcumin caused improvement of curcumin solubility and release. The 3D-construct was porous with internal interconnected pores according to SEM micrographs. Large amount of apatite was formed and proteins adsorbed on the scaffolds after immersed in physiologic buffer solution and in medium containing fetal bovine serum, respectively. Optimal concentrations of ascorbic acid and triamcinolone were used for induction of bone marrow stromal cells to become osteoblasts by expressing an enzyme marker, e.g., alkaline phosphatase. The prepared scaffolds were considered osteo-conductive and osteo-inductive. The concentrations of cisplatin and curcumin reached the IC50 of SK-ES-1 cells of osteosarcoma and MCF-7 cells of breast adenocarcinoma after 24h and 3days of release, respectively. These cancer cells were more sensitive to the combined cisplatin and curcumin than each of the drugs. Regeneration of new bone and execution of residential cancer cells in defected bone were proposed after replacing the lost bone by this established drug carrier. The assumption needs to be verified in the future using animal models.

Antioxidant and apoptotic activities of sitagliptin against hepatocellular carcinoma: An in vitro study

Background: Hepatocellular carcinoma (HCC) is the most common and aggressive type of liver cancer. Most chemotherapeutic medications nowadays imply oxidative stress leading to toxicity, which causes the necessity to find agents with better safety profiles against normal cells in addition to their anticancer activity. Sitagliptin has been shown to possess antioxidant as well as apoptotic properties by the specific suppression of dipeptidyl-peptidase 4, a glycoprotein produced in many tissues that have been thought to promote tumorigenesis and metastasis. Methods: Five groups of cell lines were included: Control (untreated HepG2 cells); cisplatin treatment HepG2 cells; sitagliptin treated HepG2 cells; combination of different concentrations of cisplatin plus sitagliptin (250 μg/mL) treated HepG2 cells, and finally, combination of different concentrations of sitagliptin plus cisplatin (25 μg/mL)-treated HepG2 cells. After an incubation period for 48 hours, the supernatants were collected to quantify the level malondialdehyde (MDA) and B-cell lymphoma-2 (BCL-2) by ELISA assay kits. Data were finally gathered and analyzed statistically. Results: Our findings indicated that sitagliptin significantly decreased the oxidative stress, particularly at high concentrations, through decreasing the MDA level. In addition, sitagliptin exhibited significant apoptotic activity against HepG2 cells through decreasing BCL-2 level. In combination with cisplatin, sitagliptin significantly potentiated the apoptotic effect and reduced the oxidative stress parameters. Conclusions: Sitagliptin showed apoptotic and antioxidant activity against HCC which may potentiate chemotherapeutic agents like cisplatin, in addition to reducing the oxidative stress against normal cells.

Kaolinite-titanium nanocomposite applied to the photodegradation of the genotoxic compound cisplatin and the terephthalic acid hydroxylation reaction

In this study, we synthesized and characterized kaolinite-titanium photocatalysts via functionalization of kaolinite with titanium (IV) isopropoxide through the sol-gel route to obtain Kaol-TiO2-400 nanocomposites for photodegradation of cisplatin. We characterized these materials by X-ray powder diffraction (XRD), Fourier-transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM). We then applied the Kaol-TiO2-400 solid in photodegradation studies after heat treatment at 400 °C and evaluated the genotoxicity of cisplatin photodegradation products. XRD results showed that the basal spacing of the Kaol solid remained unchanged at 7.14 Å, which is characteristic of pure kaolinite, indicating that only the surfaces and edges of the kaolinite lamellae were altered. The FTIR spectra of the materials displayed typical bands of Ti-OH groups and kaolinite, confirming clay surface bonding. The photodegradation studies of cisplatin by UV–visible spectroscopy and high-performance liquid chromatography (HPLC) showed that the Kaol-TiO2-400 nanocomposite had the best degradation profile, reducing the concentration of cisplatin from 15.0 mg L−1 to 5 mg L−1 within 90 min. This fast degradation profile was attributed to the well-dispersed presence of anatase on the layered clay, as observed in the XRD pattern of Kaol-TiO2-400. Additionally, the photocatalytic activity was confirmed using terephthalic acid solution as a typical model to detect the presence of hydroxyl radicals. The typical fluorescent compound 2-hydroxyterephtalyc acid was obtained within 5 min of UV light exposure. The experiment was conducted over 6 cycles, and the same profile was obtained, confirming the stability of the heterogeneous photocatalyst based on kaolinite and that no leaching of the active phase occurred during the hydroxylation reaction. Thus, the higher photodegradation profile was attributed to the presence of anatase dispersed on layered kaolinite, which was efficient in reducing the mutagenicity of cisplatin, as confirmed by biological assays.

Transcriptome Profiling of Cisplatin Resistance in Triple-negative Breast Cancer: New Insight into the Role of PI3k/Akt Pathway.

Aggressive nature of triple negative breast cancer (TNBC) is associated with poor prognosis compared with other breast cancer types. Current guidelines recommend the use of Cisplatin for the management of TNBC. However, the development of resistance to cisplatin is the primary cause of chemotherapy failure. In the present study, we aimed to develop a stable cisplatin-resistant TNBC cell line to investigate the key pathways and genes involved in cisplatin-resistant TNBC. The MDA-MB-231 cell was exposed to different concentrations of cisplatin. After 33 generations, cells showed a resistant phenotype. Then, RNA-sequencing analysis was performed in cisplatin-resistant and parent cell lines. The RNA-sequencing data was verified by quantitative PCR (qPCR). The IC50 of the resistant cell increased to 10-fold of a parental cell (p<0.001). Also, cisplatin-resistant cells show cross-resistance to other drugs, including 5- fluorouracil, paclitaxel, and doxorubicin. Resistant cells demonstrated reduced drug accumulation compared to the parental cells. Results showed there were 116 differentially expression genes (DEGs) (p<0.01). Gene ontology analysis revealed that the DEGs have several molecular functions, including binding and transporter activity. Functional annotation showed that the DEGs were enriched in the drug resistancerelated pathways, especially the PI3K-Akt signaling pathway. The most important genes identified in the protein-protein interaction network were heme oxygenase 1 (HMOX1) and TIMP metallopeptidase inhibitor 3 (TIMP3). We have identified several pathways and DEGs associated with the PI3KAkt pathway, which provides new insights into the mechanism of cisplatin resistance, and potential drug targets in TNBC.

JDHY3 Inhibits Hypopharyngeal Carcinoma Cell Proliferation and Promotes Apoptosis by Inhibiting the PI3K/AKT Pathway.

Jieduhuayu No.3 (JDHY3) is a modified Chinese herbal formula beneficial for treating hypopharyngeal carcinoma (HC), but its pharmacological mechanism is unknown. This study aimed to explore the mechanism of the herbal formula JDHY3 in inhibiting cell proliferation and promoting apoptosis in HC in vitro and in vivo. In this study, HC cells were treated with cisplatin and different concentrations of JDHY3. The apoptosis rate was detected by flow cytometry. Western blotting was used to detect the proteins related to cell proliferation and apoptosis. Afterward, the xenograft mouse model was established and treated with cisplatin and JDHY3. Mouse tumour volume was measured, and the tumour tissues were assessed by HE staining and immunohistochemistry. JDHY3 significantly inhibited the proliferation of FaDu and Detroit-562 cells. In addition, JDHY3 significantly increased the apoptosis rate of HC cells and downregulated p-PI3K and p-Akt. In addition, JDHY3 upregulated the expression of the apoptosis-promoting proteins Bax, P53, and cleaved caspase-3. In addition, the expression of the antiapoptotic protein Bcl-2 was downregulated. Coincubation with SC79 attenuated the decrease in cell proliferation induced by JDHY3, further confirming that the proapoptotic effect of JDHY3 is associated with the inhibition of PI3K/Akt pathway activation. The results of in vivo experiments showed that JDHY3 could effectively inhibit the proliferation of HC cells, and HE staining showed that JDHY3 reduced the invasion of HC cells. Immunohistochemistry showed that the expression of P53 and cleaved caspase-3 was significantly increased in the tissues of the JDHY3-treated group.

CISPLATIN - AN OVERVIEW OF ITS EFFICIENCY AND TOXICITY

Cisplatin is the first heavy metal compound that has been found to possess antineoplastic activity. It is effective in treating testicular, ovarian, head and neck, bladder, cervical, esophageal tumors, and small cell lung carcinoma. Approximately 1% of cisplatin that enters the cell interacts with DNA, forming DNA-cisplatin bonds. Both apoptosis and necrosis can be found in the same population of cells exposed to cisplatin, and the mode of cell death depends on the cisplatin concentration and metabolic state of the target cell. In the bloodstream, the platinum component of cisplatin binds to the blood's proteins (hemoglobin, albumin and transferrin), and other significant portion binds to the glutathione and other cysteine-rich biomolecules. Cisplatin impairs the mitochondrial and cell antioxidant defense system (decreases GSH, NADPH levels, GCH/GSSG ratio, and increases GSSG levels) leading to oxidative stress. There are three main mechanisms of cell resistance to cisplatin: (1) enhanced repair of cisplatin-induced DNA lesions, (2) decrease in uptake and/or increase in efflux and (3) inactivation of cisplatin intracellularly. The usage of cisplatin is limited due to its toxicity and side effects, which include neurotoxicity (numbness and tingling, paresthesia, reduced deep tendon reflexes), nephrotoxicity (renal insufficiency, hypomagnesemia), ototoxicity (tinnitus and bilateral high-frequency hearing loss), cardiotoxicity (changes in electric heart activity, congestive heart failure), gastrotoxicity (nausea, vomiting, and dyspepsia), etc. So far, there has been no effective, clinically administered, therapy for cisplatin-induced toxicity.

Synergistic Effects of New Curcumin Analog (PAC) and Cisplatin on Oral Cancer Therapy.

Oral cancer has traditionally been treated with surgery, radiotherapy, chemotherapy, or a combination of these therapies. Although cisplatin, a chemotherapy drug, can effectively kill oral cancer cells by forming DNA adducts, its clinical use is limited due to adverse effects and chemo-resistance. Therefore, there is a need to develop new, targeted anticancer drugs to complement chemotherapy, allowing for reduced cisplatin doses and minimizing adverse effects. Recent studies have shown that 3,5-Bis (4-hydroxy-3-methoxybenzylidene)-N-methyl-4-piperidine (PAC), a new curcumin analog, possesses anticancer properties and could be considered a complementary or alternative therapy. In this study, we aimed to assess the potential complementary effects of PAC in combination with cisplatin for treating oral cancer. We conducted experiments using oral cancer cell lines (Ca9-22) treated with different concentrations of cisplatin (ranging from 0.1 μM to 1 μM), either alone or in conjunction with PAC (2.5 and 5 μM). Cell growth was measured using the MTT assay, while cell cytotoxicity was evaluated using an LDH assay. Propidium iodide and annexin V staining were employed to examine the impact on cell apoptosis. Flow cytometry was used to investigate the effects of the PAC/cisplatin combination on cancer cell autophagy, oxidative stress, and DNA damage. Additionally, a Western Blot analysis was performed to assess the influence of this combination on pro-carcinogenic proteins involved in various signaling pathways. The results demonstrated that PAC enhanced the efficacy of cisplatin in a dose-dependent manner, leading to a significant inhibition of oral cancer cell proliferation. Importantly, treatment with PAC (5 μM) alongside different concentrations of cisplatin reduced the IC50 of cisplatin tenfold. Combining these two agents increased apoptosis by further inducing caspase activity. In addition, the concomitant use of PAC and cisplatin enhances oral cancer cell autophagy, ROS, and MitoSOX production. However, combined PAC with cisplatin inhibits the mitochondrial membrane potential (ΔΨm), which is a marker for cell viability. Finally, this combination further enhances the inhibition of oral cancer cell migration via the inhibition of epithelial-to-mesenchymal transition genes, such as E-cadherin. We demonstrated that the combination of PAC and cisplatin markedly enhanced oral cancer cell death by inducing apoptosis, autophagy, and oxidative stress. The data presented indicate that PAC has the potential to serve as a powerful complementary agent to cisplatin in the treatment of gingival squamous cell carcinomas.

What if amoxicillin/clavulanic acid reduces the cisplatin anticancer impact on oral cancer treatment?

Antibiotics-chemotherapeutics combination have become on the table for many cancer treatments. For this reason, we thought that further progress and development of studies to support chemotherapeutic approaches with the use of antibiotics may be beneficial in the clinical field.Cell lines (SCC-15, HTB-41, and MRC-5) were treated with 5–100 μM/ml concentrations of cisplatin (cisp) and amoxicillin/clavulanic acid (amx/cla) with combination (amx/cla-cisp) and alone in three different incubation periods. The all-cells viability was examined with WST-1 and apoptotic activity of the drugs were investigated via cell death ELISA assay kit.The cytotoxic impact of the 100 μM amx/cla-cisp combination was found to be reduced by up to 21.8%, which was significant given that the cytotoxic effect of only cisplatin therapy was 86.1%. Because our findings demonstrated that solo amx/cla therapy have almost no impact on proliferation or death, we focused on the amx/cla-cisp combination effect. It was found that the amx/cla-cisp combination has reduced the apoptotic fragment when comparing with the solely cisp-treated cells.Due to amx/cla-cisp combination on both cells but significantly on SCC-15 recovered the sole cisplatin effect, we believe that there might be a second thought when prescribing antibiotics while treating cancer patients. Not only the antibiotic's type but also the cancer type might interact to lessen the chemotherapeutic agent's impact which is clinically a dilemma to focus on.

Synergistic Effect of Anethole and Platinum Drug Cisplatin against Oral Cancer Cell Growth and Migration by Inhibiting MAPKase, Beta-Catenin, and NF-κB Pathways.

Cisplatin is a common drug used to treat patients with oral squamous cell carcinoma. However, cisplatin-induced chemoresistance poses a major challenge to its clinical application. Our recent study has shown that anethole possesses an anti-oral cancer effect. In this study, we examined the combined effect of anethole and cisplatin on oral cancer therapy. Gingival cancer cells Ca9-22 were cultured in the presence of various concentrations of cisplatin with or without anethole. The cell viability/proliferation and cytotoxicity were evaluated, respectively, by MTT, Hoechst staining, and LDH assay, while colony formation was measured by crystal violet. Oral cancer cell migration was evaluated by the scratch method. Apoptosis, caspase activity, oxidative stress, MitoSOX, and mitochondrial membrane potential (ΔΨm) levels were evaluated by flow cytometry, and the inhibition of signaling pathways was investigated by Western blot. Our results show that anethole (3 µM) potentiates cisplatin-induced inhibition of cell proliferation and decreases the ΔΨm on Ca9-22 cells. Furthermore, drug combination was found to inhibit cell migration and enhanced cisplatin cytotoxicity. The combination of anethole and cisplatin potentiates cisplatin-induced oral cancer cell apoptosis through the activation of caspase, while we also found anethole and cisplatin to enhance the cisplatin-induced generation of reactive oxygen species (ROS) and mitochondrial stress. In addition, major cancer signaling pathways were inhibited by the combination of anethole and cisplatin such as MAPKase, beta-catenin, and NF-κB pathways. This study reports that the combination of anethole and cisplatin might provide a beneficial effect in enhancing the cisplatin cancer cell-killing effect, thus lowering the associated side effects.

In vitro differences analysis in proliferation and drug sensitivity of tumor-associated fibroblasts from different regions of FDG metabolism to non-small-cell lung carcinoma

Objective: 18F-FDG PET can directly reflect the glucose metabolism level of cells from the molecular level. Tumor-associated fibroblasts, as the natural physiological barrier of tumors, support and protect the survival of non-small cell lung cancer cells. This study will further observe the differences in the supporting effects of CAFs from different regions of 18F-FDG PET metabolism on cancer cells. Methods: Biopsy tissues were collected from PET/CT patients diagnosed with NSCLC for the first time in the Department of Nuclear Medicine, The First Hospital of Shanxi Medical University. The locations of puncture were all selected in areas with high metabolism of tumor. Primary CAFs were cultured by tissue adhesion method. A549 acts as EGFR (-) tumor cells, while PC9 acts as EGFR (+) tumor cells. CAFs conditional medium was collected to induce the culture of A549 and PC9 cells, labeled as A549high/low and PC9high/low, respectively. CCK was used to measure cancer cell proliferation rate, Transwell was used to measure cancer cell invasiveness, and scratch assay was used to measure cancer cell migration. The cells were treated with cisplatin and gefitinib respectively to observe the resistance of the cells. Results: Tissues of two NSCLC patients with SUVmax were 1.25 and 11.15, respectively. The conditioned medium induced cancer cells for 48 hours. CCK (48h) results showed that SUVhigh-derived CAFs could promote the proliferation of two types of cancer cells more than SUVlow-derived CAFs. The wound healing experiment showed that A549high had completed the healing at 72h, followed by low metabolism area, and A549 was the worst. The results for PC9 cells were similar to those for A549. Transwell experiment showed that PC9high/ low showed obvious difference in invasiveness at 24 hours, and the difference was more significant at 48 hours. A549 also showed difference in invasiveness, but the difference was relatively weak. When the corresponding cells were further treated with cisplatin or gefitinib, the proliferation rate of A549high/low cells was significantly different in the gradient range of high cisplatin concentration, while the proliferation rate of PC9high/low cells was significantly different in the low-medium gefitinib concentration. Conclusion: The regions of FDG metabolism difference shown in PET support the reverse warburg effect. As a physiological barrier to tumors, CAFs is one of the key factors that promote the occurrence, metastasis and drug resistance of lung cancer. National Natural Science Foundation of China (Grant number:81971655) This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Environmental Monitoring and Ecological Risk Assessment of Platinum Complex Drugs in Wastewater Effluent: A Case Study in Qom, Iran

Background: The presence of anticancer drugs in water sources has become critical in recent years. Anticancer drugs are used in controlled amounts and conditions in medical centers. Cancerostatic platinum complexes (CPC) like cisplatin, carboplatin, and oxaliplatin are used in oncology centers to treat many cancers. Objectives: We determined the environmental risk of these compounds in municipal wastewater effluent of Qom, Iran. Methods: The LC-MS/MS technique quantified platinum complex drugs in wastewater effluent. Based on the blank laboratory method, the limit of detection (LOD) was determined as 0.009 µg/L, 0.013 µg/L, and 0.017 µg/L for oxaliplatin, carboplatin, and cisplatin, respectively. Results: The cisplatin, carboplatin, and oxaliplatin concentrations in wastewater effluent were 0.19 ± 0.098, 0.22 ± 0.094, and 0.12 ± 0.059 µg/L, respectively. Ecological risk assessment results indicated that RQsw for cisplatin, carboplatin, and oxaliplatin was 0.017, 0.013, and 0.02, respectively, showing that the platinum complex drugs had insignificant ecological exposure risk. Furthermore, ΣRQsw was estimated at &lt; 1. Conclusions: Managing cytotoxic waste from hospital oncology wards is vital for environmental pollution control. The use of other methods to remove these compounds, such as advanced oxidation processes and membrane systems, is inevitable.

Effects of baicalin on the proliferation of extranodal NK/T cell lymphoma cells and the sensitivity of the cells to the drug

Purpose: To evaluate the effect of baicalin on the proliferation of extranodal NK/T-cell lymphoma cells and the sensitivity of the cells to the drug. Methods: Cell-counting-kit (CCK) assay was used to determine cell activity, while the antagonistic, superimposed and synergistic effects of drug combinations were investigated by coefficient of drug interaction (CDI). Cell proliferation was assessed using 5-ethynyl-2’- deoxyuridine (EdU) test. Flow cytometry was carried out to determine the apoptosis of studied extranodal NK/T-cell lymphoma cells. Furthermore, western-blot was used to evaluate the expression levels of BCL2, BAX, cleaved-caspase3 and caspase3 of the cells. Results: Survival of SNK-6 and YTS cells decreased gradually in a concentration-dependent manner (r = -0.97 and -0.98). Cell viability at the same concentration diminished with time, but with increase in cisplatin concentration, cell viability at the same time point also decreased. At cisplatin concentrations of 5, 10, and 20 μmol/L, reduction in cell viability between the two groups were significantly different (p &lt; 0.001). Furthermore, the proliferation of SNK-6 and YTS cells decreased with increasing baicalin concentrations while apoptosis of the cells increased with increasing baicalin concentration. Conclusion: Baicalin inhibits the proliferation but promotes the apoptosis of SNK-6 and YTS cells. Cisplatin plus baicalin lowers cell proliferation rate and increase the apoptosis rate of both SNK-6 and YTS cells compared with baicalin or cisplatin alone. This drug combination should be further explored for its apoptosis potential in animal models.

Abstract 4925: Cannabidiol and Cannabis Sativa as a potential treatment in vitro prostate cancer cells silenced with RBBp6 and PC3 xenograft

Abstract Background: Prostate cancer is the second most frequently occurring carcinoma in males worldwide and one of the leading causes of death in men around the world. Recent studies estimate that over 1.4 million males are diagnosed with prostate cancer on an annual basis, with approximately 375 000 succumbing to the disease annually. With current treatments continuing to show severe side effects, there is a need for new treatments. In this study we looked at the effect of cannabis sativa extract, cannabidiol and cisplatin on prostate cancer cells, PC3. Methods: In addressing the above questions, we employed the MTT assay to measure the antiproliferative effect on PC3 cells following treatment with varying concentrations of Cannabis sativa extract, cisplatin and cannabidiol. xCELLigence was also used to confirm the IC50 activity in which cells were grown in a 16 well plate coated with gold and monitor cell attachment. Caspase 3/7 activity was also measured using 96 well-plate following treatment. Western-blot and qRT-PCR was also used to measure the gene expression of tumor suppressor genes, p53, Bax and Bcl2. Animal studies were employed to measure the growth of PC3-mouse derived cancer to evaluate the effect of compounds in vivo. Results: From the treatment with varying concentrations of Cannabis sativa extract, cannabidiol and cisplatin, we have observed that the three compounds induced antiproliferation of PC3 cancer cell lines through the activation of caspase 3/7 activity. We also observed induction of apoptosis in these cells following silencing of retinoblastoma binding protein 6 (RBBP6), with upregulation of p53 and bax mRNA expression, and a reduction in Bcl2 gene expression. The growth of tumors in the mouse models were reduced following treatment with cisplatin and cannabidiol. Conclusion: We demonstrated that cannabidiol is a viable therapy to treat prostate cancer cells, in combination with silencing of RBBP6. This suggests that cannabidiol rather Cannabis sativa extract may play an important role in reducing cancer progression. Citation Format: Lesetja Raymond Motadi, Zodwa E. Jantjies. Cannabidiol and Cannabis Sativa as a potential treatment in vitro prostate cancer cells silenced with RBBp6 and PC3 xenograft. [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 4925.

Abstract 5824: Human ovarian cancer cell seeding density as a mechanism of cisplatin resistance associated with expression of anti-apoptotic and autophagy proteins

Abstract Drug effects may vary depending on cancer cell density. We investigated discrepancies in phenotypes for high density and low-density cultured cancer cells and elucidated relevant variables that influence them. We assessed chemosensitivity as a function of tumor cell density by plating TOV-21G and OVMANA ovarian cancer cell lines in 96-well plates at various seeding densities per well and treated them with various concentrations of cisplatin. CellTiterGlo assays were performed at 72 hours of treatment to determine cisplatin dose response curves. We used ethidium homodimer fluorescence staining and imaging to quantify extent of cell death at differing cell densities of TOV-21G and OVMANA cells treated with cisplatin for 48 and 72hours in 48-well plates. We performed western blot analyses of apoptotic and autophagy proteins from TOV-21G and OVMANA cells that were seeded at high and low cell densities in 24-wellplates. TOV-21G and OVMANA cells plated at higher cell seeding densities displayed less cisplatin sensitivity than TOV-21G and OVMANA cells plated at lower seeding densities, as shown by cisplatin dose response curves. Ethidium homodimer staining of TOV-21G and OVMANA cells treated with cisplatin revealed that cells seeded at lower densities experience a greater amount of cell death that cells seeded at higher densities. Western blot analyses showed that high density TOV-21G cells may express greater amounts of BCL2 and c-FLIP and lower amounts of p62 and NOXA compared to low density cells when normalized for protein expression. Similarly, high density OVMANA cells express greater amounts of c-FLIP andBCL2 and lower amounts of NOXA compared to low density cells when normalized for protein expression. Our experiments reveal that human ovarian cancer cells display different phenotypic changes depending on cell density, causing higher density cells to require increased cisplatin to achieve the same level of effect seen in lower density cells. Ovarian cancer cell resistance to cisplatin is associated with alterations in autophagy and anti-apoptotic protein expression at high cell seeding density. These alterations may also affect chemosensitivity in vivo in settings of differential tumor density, such as single circulating tumor cells compared to clumps of tumor cells, and other settings, such as metastasis. Citation Format: Thomas W. Brown, Ujwal Punyamurtula, Wafik S. El-Deiry. Human ovarian cancer cell seeding density as a mechanism of cisplatin resistance associated with expression of anti-apoptotic and autophagy proteins. [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 5824.

Abstract 4936: Activity of carfilzomib in head and neck cancer cell lines in vitro

Abstract Background: Carfilzomib (CF) is a 2nd generation proteasome inhibitor with activity in hematologic cancer, but its study in solid tumors like squamous cell carcinoma of the head and neck (SCCHN) is limited. We investigated its antitumor activity in three SCCHN cell lines. Methods: Cells: We studied two human tongue cell lines (CAL27; SCC-25) and one pharynx line (FaDu); all HPV-negative and p53-mutant. MTT assays: We utilized MTT assays to evaluate cytotoxicity of CF, cisplatin, and their combination. The IC50 of CF and Combination Index (CI) of varying concentrations of combined CF and cisplatin were calculated. Clonogenic assays: Cells were seeded at low density and after 24 hours, cells were treated with varying concentrations of CF. Plates were incubated for 10 days. The cells were fixed and stained with crystal violet dye and colonies counted. Wound Healing assays: After cells reached 100% confluence, a scratch was created using a 200 µL micropipette. Cells were treated with CF and images were acquired at 0, 24, 48, and 72 hours post-scratch. Results: MTT assays: IC50 values as a function of exposure lengths are shown in Table 1. CAL27 was the most sensitive while SCC-25 was the most resistant. In all cell lines, the results of combined CF and cisplatin were complex: at lower doses there was evidence of antagonism, while at higher doses there was synergy. Clonogenic assays: CF significantly inhibited clonogenicity compared to DMSO control (P≤ 0.05). The surviving fraction for CF 2.5, 5, and 10 nM were 0.411. 0.151, 0.051, and 0.5, 0.339, 0.102 in CAL27 and SCC-25 cell lines, respectively. Wound Healing assays: Wound closure % after treatment with CF5 was significantly different from DMSO control (P≤0.05). Wound closure % for CF5 were 30.97, 56.75, and 28.03% compared to 94.65, 75.88, and 42.05% for control in CAL27, FaDu, and SCC-25 cell lines, respectively. Conclusion: CF exhibits antitumor effects in SCCHN in vitro. Our findings support further investigation of CF in combination with conventional agents in vitro and in vivo. IC50 values for carfilzomib in head and neck cancer cell lines Cell line 24 hours 48 hours 72 hours CAL27 18.02 (11.76- 28.22) 9.364 (7.189- 12.45) 6.401 (5.579- 7.373) FaDu 57.34 (41.98-84.90) 16.22 (13.438- 19.446) 8.237 (7.260- 9.365) SCC25 77.78 (67.18- 91.30) 12.87 (11.36- 14.64) 11.49 (10.51- 12.58) Citation Format: Abdullah S. Eldaly, Douglas B. Stairs, Mitchell Machtay. Activity of carfilzomib in head and neck cancer cell lines in vitro. [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 4936.

Abstract 2852: Targeting NPEPPS overcomes cisplatin resistance in patient-derived bladder cancer tumoroids

Abstract Introduction and Objectives: Cisplatin-based neoadjuvant chemotherapy (NAC) is the recommended treatment for muscle-invasive bladder cancer (MIBC) patients. Patients with a pathological complete response after NAC (pCR), have a good 5-yr OS of 80%. However, due to NAC resistance, only 25% of patients achieve a pCR. Our aim is to increase the number of patients reaching a pCR by overcoming cisplatin resistance. Previously we identified NPEPPS as a key player in cisplatin resistance that prevents cisplatin uptake via volume-regulated anion channels (VRAC) in cisplatin-resistant cell lines. Studies have demonstrated that patient-derived bladder cancer tumoroids recapitulate the originating tumor at the molecular level, presenting a model system to study chemotherapy resistance ex vivo. Here, we used MIBC tumoroids to functionally interrogate NPEPPS’ role in cisplatin resistance. Material and Methods: Ex vivo interrogation of NPEPPS function in cisplatin resistance was performed in tumoroid cultures generated from MIBC patients (N=10). Tumor tissue was obtained either at transurethral resection of bladder tumor (TURBT, N=5) or RC (N=5), and clinical resistance to NAC was defined as ypT≥2, nodal positive disease at RC or progression during NAC. Molecular characterization of tumor specimens and corresponding tumoroids was performed by bladder cancer-specific SNaPshot mutation analysis, copy-number aberrations analysis, immunohistochemistry (IHC), and hematoxylin &amp; eosinophilic staining (HE). The effect of NPEPPS on cisplatin uptake was investigated using cisplatin treatment combined with the pharmacological NPEPPS-inhibitor tosedostat, shRNA-mediated NPEPPS knock-down or lentiviral NPEPPS over-e xpression followed by cytometry by time of flight (CyTOF) analysis. CellTiter-Glo cell viability assays and caspase 3/7 apoptosis assays were used to investigate whether NPEPPS inhibition overcomes cisplatin resistance ex vivo. Results: Five out of ten patients received NAC, all whom had tumors that were clinically resistant to NAC. Tumoroid cultures from MIBC patients unresponsive to NAC showed resistance to serum concentrations of cisplatin as well as increased NPEPPS mRNA expression. Meanwhile, molecular characterization of tumor and tumoroid pairs confirmed that patient-specific tumor traits were maintained in tumoroids. Pharmacological NPEPPS inhibition or NPEPPS depletion by shRNA-mediated knock-down increased cisplatin influx into the cell and resensitized NAC resistant tumoroids to serum concentrations of cisplatin. Furthermore, NPEPPS overexpression increased cisplatin resistance in cisplatin-sensitive tumoroids. Conclusion: We confirm that NPEPPS is associated with cisplatin-resistance in an ex vivo MIBC tumoroid model. These findings have potential for rapid translation into the clinic and invite trials investigating tosedostat to overcome chemoresistance. Citation Format: Mathijs P. Scholtes, Maryam Akbarzadeh, Shahla Romal, Miranda van Dijk, Tsung Wai Kan, Tokameh Mahmoudi, Tahlita C. Zuiverloon. Targeting NPEPPS overcomes cisplatin resistance in patient-derived bladder cancer tumoroids [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 2852.