Exploring the Role of Mucin 13 in Hepatocellular Carcinoma

Background: Hepatocellular carcinoma (HCC) has a poor prognosis due to ineffective therapeutic modality and lack of early diagnostic marker. Accumulating studies have shown that elevated expression of mucin 13 as potential oncogene and predictive biomarker for various cancer. However, very little is known about its expression and function for development and progression of HCC. Objective: To investigate mucin 13 expression in chemically induced hepatocellular carcinoma model. Methodology: Diethyl nitrosamine (DEN) and 2-Acetylaminofluorene (2-AAF) induced method was employed for the development of hepatocellular carcinoma in Male Wistar rats. Serum and tissues were collected at regular intervals of time and routinely validated for liver cancer stages. Immunohistochemistry and in situ hybridization were performed on formalin-fixed, paraffin-embedded tissues. Molecular docking studies were performed to study the interaction of mucin 13 and DEN. Results: Our results demonstrate hepatocellular adenoma as observed by histopathological analysis. Biochemical analysis showed a progressive increase in the levels of serum ALT, AST, and ALP, suggesting the development and progression of hepatocellular damage. Notably, mucin 13 expression gradually elevated during consecutive stages of hepatocellular carcinoma. Interestingly, an increase in nuclear localization of mucin 13 was observed in the treated group as compared to control group. In situ hybridization analysis showed that a decrease in miR-132 and miR-145, which are inversely related with mucin 13 expression. Moreover, DEN efficiently binds mucin 13 with high affinity and thus stabilizes it as demonstrated by molecular docking analysis. Conclusion: These results suggest that mucin 13 expression is closely associated with hepatocarcinogenesis and could serve as a predictive candidate biomarker for HCC

Exosome-Based Smart Drug Delivery for Cancer Treatment

Advances in nanoscale materials have become indispensable for targeted drug delivery, early detection, and personalized approaches for cancer treatment. Among various nanoscale materials investigated, exosomes hold significant promise in drug delivery. Exosomes are nanoscale vesicles that are usually 30–150 nm in size and produced by cells for intercellular communication. Due to their unique composition and inherent tumor-targeting capacity, these particles are well suited for tumor-specific delivery systems. This chapter discusses exosome isolation, therapeutic loading methods, key roles of exosomes in the tumor microenvironment, current applications of exosomes in drug delivery, and possible clinical implications

Cucurbitacin D ameliorates benzo[a]pyrene induced liver injury via Activation of Nrf2 Antioxidant pathway

Background: Co-morbidity variables, such as smoking, are strongly linked to the development and progression of liver cancer. Further, benzo[a]pyrene, a major component of tobacco smoke, is highly carcinogenic and triggers liver damage. Cucurbitacin, a kind of triterpene, has a wide range of biological properties, including antioxidant, anti-inflammatory, and anti-cancer effects. However, the precise mechanism of its hepatoprotective effects is obscure. Objective: The aim of this study is to investigate the cytoprotective effects of novel analog of cucurbitacin, cucurbitacin D, against benzo[a]pyrene-induced liver injury in HepG2 cells. Method: The cytoprotective efficacy of cucurbitacin D against benzo[a]pyrene-induced liver damage was studied using proliferation, clonogenicity, migration, invasion, Western blotting, and qPCR analysis. The levels of intracellular reactive oxygen species (ROS) in liver cells was measured using the DCFDA assay. Results: In human HepG2 cells, functional experiments revealed that cucurbitacin D has cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene. The mitigation of ROS observed by fluorimeter and fluorescence microscopy suggested that this protective effect was likely due to cucurbitacin D\u27s antioxidant property. Additional research is ongoing to identify the effect of cucurbitacin D on oxidative stress markers by using qPCR and western blotting techniques. Overall, these findings showed that cucurbitacin D diminishes benzo[a]pyrene-induced liver injury via its antioxidant activity. Conclusion: These findings show that cucurbitacin D has hepatoprotective properties against benzo[a]pyrene-induced liver injury, making it an attractive food supplement ingredient

Hepatoprotective role of Cucurbitacin D on benzo[a]pyrene induced liver injury

Background: Epidemiological findings show the strong correlation of co-morbidity factors including smoking with the development and progression of liver cancer. Moreover, benzo[a]pyrene, a main component of tobacco smoke, is extremely carcinogenic and contributes to liver injury as well. Cucurbitacin, chemically classified as triterpenes, have shown diverse biological activities including potent antioxidant, anti-inflammatory and anti-cancer activities. However, their hepatoprotective activities are not completely understood. Objective: In the present study, we investigated the cytoprotective activity of novel analog of cucurbitacin, cucurbitacin D, against benzo[a]pyrene-induced liver injury in human HepG2 cells. Method: Proliferation, clonogenicity, migration, invasion, Western blotting and qPCR analysis were conducted to investigate the cytoprotective effect of cucurbitacin D against benzo[a]pyrene induced liver damage. DCFDA assay was performed to analyze intracellular reactive oxygen species (ROS) level in liver cells. Results: Functional assays showed that cucurbitacin D exhibited cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene in human HepG2 cells. This protective effect was likely associated with antioxidant potential of cucurbitacin D, as evidenced by the attenuation of ROS observed by fluorimeter and fluorescence microscopy. Further study is ongoing to examine the effect of cucurbitacin D on oxidative stress markers by employing western blotting and qPCR techniques. Collectively, these results exhibited that cucurbitacin D alleviate benzo[a]pyreneinduced liver injury through its antioxidant effects. Conclusion: These results have demonstrated hepatoprotective effects of cucurbitacin D against benzo[a]pyrene-induced liver damage, rendering it as an effective potential ingredient in food supplements

Pharmacognostical and Phytochemical Evaluation of a Unani Polyherbal Formulation: Dawa ul Kurkum by HPTLC

Background: Dawa ul Kurkum (Duk) is a widely used Unani formulation. It consists of seven plant herbs, including stigmas of Crocus sativus L., rhizomes of Nardostachys jatamansi (D.Don) DC., the bark of Cinnamomum cassia (L.) J. Presl., shoot of Cymbopogon jwarancusa (Jones ex Roxb.) Schult., the resin of Commiphora wightii (Arn.) Bhandari, roots of Saussurea lappa (Decne.) Sch.Bip., and bark of Cinnamomum zeylanicum Blume. However, no study has been previously conducted to characterize this formulation. Thus, the present study was designed to carry out the pharmacognostic and phytochemical characterization of Duk. Methods: Duk was prepared following the protocols in Bayaz e Kabeer and The National Formulary of Unani Medicine Part-I. The characterization included organoleptic properties, fluorescence analysis, preliminary phytochemical screening, antioxidant activity, and active constituent profiling using HPTLC. Results: Evaluation of Duk showed the presence of carbohydrates, flavonoids, quinones, glycosides, cardiac glycosides, terpenoids, phenols, coumarin, steroids, and phytosterols. The total phenolic and flavonoid content was 5.75 ± 0.23 mg GAE/g and 10 ± 0.18 mg QUE/g, respectively. HPTLC of Duk showed the presence of p-coumaric acid, cinnamaldehyde, citral, crocin, isovaleric acid, guggulsterone, and dehydrocostus lactone. Conclusions: Our findings supported the use of Duk as a conventional medicine, and these results could be used as a reference for the standardization of Duk

Tannic Acid Exhibits Antiangiogenesis Activity in Nonsmall-Cell Lung Cancer Cells

Nonsmall-cell lung cancer (NSCLC) is the most common type of lung cancer, with a dismal prognosis. NSCLC is a highly vascularized tumor, and chemotherapy is often hampered by the development of angiogenesis. Therefore, suppression of angiogenesis is considered a potential treatment approach. Tannic acid (TA), a natural polyphenol, has been demonstrated to have anticancer properties in a variety of cancers; however, its angiogenic properties have yet to be studied. Hence, in the current study, we investigated the antiproliferative and antiangiogenic effects of TA on NSCLC cells. The (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) (MTS) assay revealed that TA induced a dose- and time-dependent decrease in the proliferation of A549 and H1299 cells. However, TA had no significant toxicity effects on human bronchial epithelial cells. Clonogenicity assay revealed that TA suppressed colony formation ability in NSCLC cells in a dose-dependent manner. The anti-invasiveness and antimigratory potential of TA were confirmed by Matrigel and Boyden chamber studies, respectively. Importantly, TA also decreased the ability of human umbilical vein endothelial cells (HUVEC) to form tube-like networks, demonstrating its antiangiogenic properties. Extracellular vascular endothelial growth factor (VEGF) release was reduced in TA-treated cells compared to that in control cells, as measured by the enzyme-linked immunosorbent assay (ELISA). Overall, these results demonstrate that TA can induce antiproliferative and antiangiogenic effects against NSCLC

Cucurbitacin D suppresses benzo[a]pyrene-induced liver injury by modulating Nrf2 signaling

Background: Accumulating studies have shown strong correlation of HCC and co-morbidity factors including smoking. Tobacco smoke contains benzo[a]pyrene, which is extremely carcinogenic and contributes to liver damage. Cucurbitacin, a triterpene, has a wide range of biological activities, including antioxidant, anti-inflammatory, and anti-cancer properties. However, their hepatoprotective effects remain poorly understood. In the current study, we examined the hepatoprotective activity of cucurbitacin D, a novel analog of cucurbitacin, against benzo[a]pyrene-induced liver injury in human HepG2 cells. Method: To investigate the hepatoprotective effect of cucurbitacin D against benzo[a]pyrene-induced liver damage, proliferation, clonogenicity, migration, invasion, Western blotting, and qPCR analyses were performed. The DCFDA assay was performed to determine the level of intracellular reactive oxygen species (ROS) in liver cells. Results: Functional assays showed that cucurbitacin D exhibited cytoprotective effects against dose-dependent growth inhibition by benzo[a]pyrene in human HepG2 cells. This protective effect was likely associated with antioxidant potential of cucurbitacin D, as evidenced by the attenuation of ROS observed by fluorimeter and fluorescence microscopy. Western blotting analysis demonstrated Cucurbitacin D targets Nrf-2 signaling pathway and associated effector proteins including HO-1 and LC3A in protecting liver cells against benzo[a]pyrene induce oxidative damage. Further studies are underway to understand the underlying molecular mechanism of action. Conclusion: These findings demonstrate the hepatoprotective effects of cucurbitacin D against benzo[a]pyrene-induced liver damage, making it a promising ingredient for nutritional supplements

Therapeutic efficacy of ormeloxifene against hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) is the most common primary liver cancer and leading cause of cancer related deaths worldwide. Severe toxicity and drug resistance to available chemotherapeutic agents display ineffective clinical response. Therefore, drug repurposing is gaining attention owing to their known biological activities and excellent safety profiles. Ormeloxifene (ORM), non-steroidal, selective estrogen receptor modulator (SERM), and exhibit diverse pharmacological activities. The aim of this study is to assess the therapeutic activity of ORM and to investigate the underlying molecular mechanism against hepatocellular carcinoma. Objective: To investigate the therapeutic activity of ormeloxifene in human hepatocellular carcinoma cells. Methodology: MTT and colony formation assays were performed in SK-Hep-1, Hep3B and C3A cells. In vitro functional assays were carried out for investigating effect of ORM on migration and invasion abilities of HCC cells using Boyden chamber and Matrigel assays respectively. Results: Functional analysis revealed that ORM treatment led to suppression of proliferation and colony formation in human hepatocellular carcinoma cells in dose and time-dependent manner compared to vehicle treated group. ORM treatment, as shown by wound healing and Matrigel invasion assay, respectively, suppresses the migration and invasion of human hepatocellular carcinoma cells. Further, experiments are underway to determine the effect of ORM on EMT markers using western blotting and qPCR techniques. Conclusion: Taken together, ORM exhibited potent anticancer effects against HCC and could be further explored as a novel therapeutic modality for the treatment of HCC

Studying the molecular mechanism of hepatocellular carcinoma

Background: Hepatocellular carcinoma (HCC) has a poor prognosis due to ineffective therapeutic modality and lack of early diagnostic marker. Accumulating studies have shown that elevated expression of mucin 13 as potential oncogene and predictive biomarker for various cancer. However, very little is known about its expression and function for development and progression of HCC. Objective: To investigate mucin 13 expression in chemically induced hepatocellular carcinoma model. Methodology: Diethyl nitrosamine (DEN) and 2-Acetylaminofluorene (2-AAF) induced method was employed for the development of hepatocellular carcinoma in Male Wistar rats. Serum and tissues were collected at regular intervals of time and routinely validated for liver cancer stages. Immunohistochemistry and in situ hybridization were performed on formalin-fixed, paraffin-embedded tissues. Molecular docking studies were performed to study the interaction of mucin 13 and DEN. Results: Our results demonstrate hepatocellular adenoma as observed by histopathological analysis. Biochemical analysis showed a progressive increase in the levels of serum ALT, AST, and ALP, suggesting the development and progression of hepatocellular damage. Notably, mucin 13 expression gradually elevated during consecutive stages of hepatocellular carcinoma. Interestingly, an increase in nuclear localization of mucin 13 was observed in the treated group as compared to control group. In situ hybridization analysis showed that a decrease in miR-132 and miR-145, which are inversely related with mucin 13 expression. Moreover, DEN efficiently binds mucin 13 with high affinity and thus stabilize it as demonstrated by molecular docking analysis. Conclusion: These results suggest that mucin 13 expression is closely associated with hepatocarcinogenesis and could serve as a predictive candidate biomarker for HCC

Pharmacological restoration of PKD1: A novel strategy for prostate cancer therapy

Background: Prostate cancer has poor prognosis owing to late diagnosis and ineffective multimodal clinical treatment. Extensive efforts are ongoing to establish methods that can resolve the expression of genes implicated in disease development and treatment. Previously, we reported that Protein Kinase D1 (PKD1), a serine threonine kinase, controls a number of tumor suppressor functions including cell aggregation, cell motility, cell proliferation, and cell invasion. Thus, PKD1 is considered as an emerging therapeutic target for prostate cancer treatment. Objective: To investigate the restoration of PKD1 by a pharmacological modulator ormeloxifene, which showed well-defined PK/PD and safety profiles in humans. Methods: Proliferation, clonogenicity, migration, invasion, western blotting and qPCR analysis were performed to investigate the anticancer effect of ORM, docetaxel and/or their combination on PKD1 and related signaling mechanisms in prostate cancer. Results: ORM treatment inhibited cell proliferation, invasion, migration and colony formation abilities of prostate cancer cells in a dose-dependent manner compared to vehicle treated group. ORM treatment selectively induces the expression of PKD1 both at mRNA and protein levels in C4-2 cells. Moreover, our results have also shown that ORM effectively attenuates MTA1 expression in prostate cancer cells. MTA1 physically interact and shown to have inverse relationship with PKD1. In addition, we observed that ORM treatment enhances the therapeutic efficacy of docetaxel in C4-2 cells. Our results also indicate that ORM treatment potentiate the effects of docetaxel as determined by MTS and colony formation assays. Conclusion: These results suggest that ORM exhibit potent anticancer activity via restoration of PKD1 in prostate cancer