Early Diagnosis Of Liver Cancer Research Articles

A DNA-peptide probe coupled mass spectrometry-based method for the detection of LncRNA in hepatocyte and serum samples

Liver cancer, a prominent contributor to cancer-related deaths, necessitates timely identification for successful intervention. In recent years, long non-coding RNAs (lncRNAs) have gained attention as potential biomarkers for liver cancer, and accurate quantification of lncRNA is essential for early diagnosis of liver cancer. However, existing methods for lncRNA detection rely on amplification and labeling techniques, resulting in semi-quantitative or qualitative outcomes. DNA-peptide technology offers a promising alternative. It has successfully translated nucleic acid quantification into peptide quantification, particularly in the context of miRNA analysis. However, the intricate structure of long-chain lncRNAs presents a challenge not encountered with miRNAs, making the design and acquisition of specific DNA-peptide probes for direct quantification more difficult. In this study, a strategy for lncRNA quantification was developed using a DNA-peptide probe coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based quasi-targeted proteomics. The inherent structural complexity of long-chain lncRNAs was overcome by designing DNA-peptide probes based on their unique sequence segments, enabling direct quantification without the need for amplification. Highly upregulated in liver cancer (HULC) was identified as a target lncRNA through bioinformatics analysis using GEO2R online software. Subsequently, the target lncRNA (i.e., HULC), which had been biotinylated and bound to streptavidin agarose beforehand, was hybridized with the probe. Following trypsin digestion, the reporter peptide was released and quantified using LC-MS/MS. Finally, this method was applied to analyze HULC levels in liver cancer cell lines and serum samples, enabling quantitative assessment and early diagnosis. The results demonstrated that liver cancer diagnostics could be enhanced through the integration of mass spectrometry and DNA-peptide probe technology for lncRNA quantification.

Dual-reporter fluorescent probe for precise identification of liver cancer by sequentially responding to carboxylesterase and polarity

Early treatment significantly improves the survival rate of liver cancer patients, so the development of early diagnostic methods for liver cancer is urgent. Liver cancer can develop from viral hepatitis, alcoholic liver, and fatty liver, thus making the above diseases share common features such as elevated viscosity, reactive oxygen species, and reactive nitrogen species. Therefore, accurate differentiation between other liver diseases and liver cancer is both a paramount practical need and challenging. Numerous fluorescent probes have been reported for the diagnosis of liver cancer by detecting a single biomarker, but these probes lack specificity for liver cancer in complex biological systems. Obviously, using multiple liver cancer biomarkers as the basis for judgment can dramatically improve diagnostic accuracy. Herein, we report the first fluorescent probe, LD-TCE, that sequentially detects carboxylesterase (CE) and lipid droplet polarity in liver cancer cells with high sensitivity and selectivity, with linear detection of CE in the range of 0–6 U/mL and a 65-fold fluorescence enhancement in response to polarity. The probe first reacts with CE and releases weak fluorescence, which is then dramatically enhanced due to the decrease in lipid droplet polarity in liver cancer cells. This approach allows the probe to enable specific imaging of liver cancer with higher contrast and accuracy. The probe successfully achieved the screening of liver cancer cells and the precise identification of liver cancer in mice. More importantly, it is not disturbed by liver fibrosis, which is a common pathological feature of many liver diseases. We believe that the LD-TCE is expected to be a powerful tool for early diagnosis of liver cancer.

Early monitoring values of oncogenic signalling molecules for hepatocellular carcinoma.

The prevention and early diagnosis of liver cancer remains a global medical challenge. During the malignant transformation of hepatocytes, a variety of oncogenic cellular signalling molecules, such as novel high mobility group-Box 3, angiopoietin-2, Golgi protein 73, glypican-3, Wnt3a (a signalling molecule in the Wnt/β-catenin pathway), and secretory clusterin, can be expressed and secreted into the blood. These signalling molecules are derived from different signalling pathways and may not only participate in the malignant transformation of hepatocytes but also become early diagnostic indicators of hepatocarcinogenesis or specific targeted molecules for hepatocellular carcinoma therapy. This article reviews recent progress in the study of several signalling molecules as sensitive biomarkers for monitoring hepatocarcinogenesis.

Utilization of Biopsy-Guided CT Scan in Diagnosing Liver Cancer: A Case Study

Introduction: Liver cancer is one of the most common types of cancer in Indonesia and has a high mortality rate. Early diagnosis of liver cancer is very important to increase the patient's chances of recovery. Biopsy-guided CT scan is an effective method for diagnosing liver cancer.
 Case presentation: We report the case of a 55 year old man with a history of chronic hepatitis B who presented with complaints of right upper abdominal pain and weight loss. Physical examination revealed hepatomegaly and ascites. Investigations, including abdominal ultrasound and liver function tests, showed a mass in the liver. CT scan of the abdomen with contrast showed a hypodense mass in the right hepatic lobe. A CT-guided liver biopsy was performed and the histopathological diagnosis was hepatocellular carcinoma (HCC). The patient then underwent partial resection hepatectomy and chemotherapy.
 Conclusion: Biopsy-guided CT scan is a valuable tool for the diagnosis of HCC in patients with chronic hepatitis B.

A compact microfluidic laser-induced fluorescence immunoassay system using avalanche photodiode for rapid detection of alpha-fetoprotein.

Alpha-fetoprotein (AFP), commonly employed for early diagnosis of liver cancer, serves as a biomarker for cancer screening and diagnosis. Combining the high sensitivity and specificity of fluorescence immunoassay (FIA), developing a low-cost and efficient immunoassay system for AFP detection holds significant importance in disease diagnosis. In this work, we developed a miniaturized oblique laser-induced fluorescence (LIF) immunoassay system, coupled with a microfluidic PMMA/paper hybrid chip, for rapid detection of AFP. The system employed an avalanche photodiode (APD) as the detector, and implemented multi-level filtering in the excitation light channel using the dichroic mirror and optical trap. At first, we employed the Savitzky-Golay filter and baseline off-set elimination methods to denoise and normalize the original data. Then the cutoff frequency of the low-pass filter and the reverse voltage of the APD were optimized to enhance the detection sensitivity of the system. Furthermore, the effect of laser power on the fluorescence excitation efficiency was investigated, and the sampling time during the scanning process was optimized. Finally, a four-parameter logistic (4PL) model was utilized to establish the concentration-response equation for AFP. The system was capable of detecting concentrations of AFP standard solution within the range of 1-500 ng/mL, with a detection limit of 0.8 ng/mL. The entire immunoassay process could be completed within 15 min. It has an excellent potential for applications in low-cost portable diagnostic instruments for the rapid detection of biomarkers.

Factors affecting the compliance of hepatocellular carcinoma screening among high-risk populations: A systematic review and meta-analysis.

Hepatocellular carcinoma (HCC) is the sixth most common cancer worldwide and the third leading cause of cancer mortality. HCC has high morbidity, high mortality, and low survival rates. Screening is one of the most significant methods of lowering incidence and death while also increasing survival. The aim of this study was to identify the facilitators and barriers to participation in HCC screening among high-risk populations. A comprehensive and systematic search was undertaken in PubMed, Web of Science, MEDLINE, EMBACE, EBSCOhost and the Cochrane Library. A combination of synonyms of the keywords including HCC, screening, factors and adherence were used for searching. Studies addressing the facilitators and barriers to HCC screening compliance in at-risk individuals were included. Data were synthesized using Review Manager version 5.4. A random/fixed effects model meta-analysis was performed to estimate the pooled data and expressed with odds ratio (OR) and 95% confidence interval (CI). A total of seven articles met the inclusion criteria. Qualitative (n=1) and quantitative (n=6) studies using various types of surgery were conducted. The most commonly mentioned barriers were insufficient knowledge and awareness of HCC screening, unawareness of the necessity for early detection of HCC and lack of physician recommendation. A meta-analysis of seven studies showed that individuals with a family history of HCC increased screening uptake by nearly three times (OR: 2.69, 95% CI: 1.93, 3.75). Other most frequently reported facilitators include age, education level, and perceived risk etal. Many barriers to HCC screening were found. Meanwhile, this review points out that improving the awareness of high-risk populations toward HCC screening is expected to enhance compliance, thereby promoting early diagnosis of liver cancer, reducing mortality, and alleviating the burden of HCC.

Methylene blue based near-infrared fluorogenic probe for lighting up esterase activity and its application in biological imaging

In this work, a light-up near-infrared (NIR) fluorescence probe based on methylene blue was developed for the determination of esterase. Probe MB-Ace emits a significant NIR fluorescence at 691 nm while incubated with esterase, which facilitates us to accurately detect the activity of esterase in vitro and vivo. MB-Ace showed high sensitivity (with a detection limit of 5.2 ng/mL) and selectivity to esterase under the interference of various biomolecules. The results of biological imaging experiments showed that MB-Ace had cell membrane permeability and could be used to monitor esterase activity in Hep G2 cells. At the same time, MB-Ace also showed excellent imaging ability for esterase activity in a mice model. The above results reflect the application value of MB-Ace in the early diagnosis of liver cancer and image-guided tumor resection.

MS-FANet: Multi-scale feature attention network for liver tumor segmentation

Accurate segmentation of liver tumors is a prerequisite for early diagnosis of liver cancer. Segmentation networks extract features continuously at the same scale, which cannot adapt to the variation of liver tumor volume in computed tomography (CT). Hence, a multi-scale feature attention network (MS-FANet) for liver tumor segmentation is proposed in this paper. The novel residual attention (RA) block and multi-scale atrous downsampling (MAD) are introduced in the encoder of MS-FANet to sufficiently learn variable tumor features and extract tumor features at different scales simultaneously. The dual-path feature (DF) filter and dense upsampling (DU) are introduced in the feature reduction process to reduce effective features for the accurate segmentation of liver tumors. On the public LiTS dataset and 3DIRCADb dataset, MS-FANet achieved 74.2% and 78.0% of average Dice, respectively, outperforming most state-of-the-art networks, this strongly proves the excellent liver tumor segmentation performance and the ability to learn features at different scales.

FRI-323 - Exploration of a holistic management procedure for liver cancer surveillance to improve the early diagnosis of liver cancer in Chinese population

FRI-323 - Exploration of a holistic management procedure for liver cancer surveillance to improve the early diagnosis of liver cancer in Chinese population

Highly sensitive paper-based immunoassay: Combining nanomaterials modified cellulose with covalent and oriented immobilization of antibody

Among the analytical tools, paper-based analytical devices (PADs) have become a leading alternative for point-of care testing (POCT). In this study, PADs were fabricated using an office laser printer. Then, the paper zone was modified with graphene oxide (GO) and pyrene derivatives, which provide a sufficient amount of carboxylic groups for conjugating antibodies. At an optimal pH, antibodies were covalently bound onto carboxylated cellulose surface in an oriented manner through a two-step strategy: electrostatic adsorption was followed by EDC/NHS coupling. α-fetoprotein (AFP) as a detection model, we compared with cellulose powder modified and unmodified paper zone. The results showed the color intensity and color uniformity on GO modified paper was improved. The activity of immobilized antibodies on GO/1-pyrenebutyric acid (GO/PBA) modified was three times higher than that of GO modified and about 1.8-fold higher than that of GO/1-pyrenecarboxylic acid (GO/PCA) modified. The GO/PBA modified paper-based immunoassay has enhanced sensitivity and low detection limit. A linear correlation between color intensity and concentration of AFP in the range of 0.01～16.5 ng mL−1 with a detection limit of 9.0 pg mL−1 were achieved, respectively. The obtained results point towards rapid, sensitive, and specific early diagnosis of liver cancer at the point of care and other low-resource settings.

Identification of Liver Cancer Driver Mutations from COSMIC Data

Background: Liver cancer accounts for more than 700,000 deaths each year making it the third leading cause of cancer-related deaths worldwide. Late diagnosis of the disease is the reason behind most deaths. Driver mutations are genetic alterations in tumor cells, which are responsible for the development of liver cancer; therefore, the identification of genetic biomarkers is necessary for the prediction and early diagnosis of liver cancer. Objectives: The main objective of this study is to identify pathogenic alleles that may act as potential biomarkers for the prediction of liver cancer. It also identifies the role of novel genes in liver cancer that are not known to cause the disease. Methods: The mutation data of non-coding variants were downloaded from the catalogue of somatic mutations in cancer (COSMIC) databases. Different bioinformatics tools were, then, used to retrieve mutations in liver cancer. The genetic alterations in hepatocellular carcinoma (HCC) were analyzed. Results: The present study successfully identified pathogenic alleles (consistent mutations) along with a set of novel genes that might be involved in the development of liver cancer. It identified non-coding mutations near human genes and transcription factor binding sites of HepG2 cells. This study also identified mutations near the genes that are involved in the Ras/MAFK signaling pathway of the Hepatitis B virus. Conclusions: The pathogenic alleles identified in this study may provide targeted therapy for the treatment of liver cancer. The identification of novel genes may help to understand the progression of liver cancer at the molecular level. The identified driver mutations may act as potential biomarkers and therapeutic targets for early prediction and treatment of liver cancer.

Evaluation and Application of Drug Resistance by Biomarkers in the Clinical Treatment of Liver Cancer.

Liver cancer is one of the most lethal cancers in the world, mainly owing to the lack of effective means for early monitoring and treatment. Accordingly, there is considerable research interest in various clinically applicable methods for addressing these unmet needs. At present, the most commonly used biomarker for the early diagnosis of liver cancer is alpha-fetoprotein (AFP), but AFP is sensitive to interference from other factors and cannot really be used as the basis for determining liver cancer. Treatment options in addition to liver surgery (resection, transplantation) include radiation therapy, chemotherapy, and targeted therapy. However, even more expensive targeted drug therapies have a limited impact on the clinical outcome of liver cancer. One of the big reasons is the rapid emergence of drug resistance. Therefore, in addition to finding effective biomarkers for early diagnosis, an important focus of current discussions is on how to effectively adjust and select drug strategies and guidelines for the treatment of liver cancer patients. In this review, we bring this thought process to the drug resistance problem faced by different treatment strategies, approaching it from the perspective of gene expression and molecular biology and the possibility of finding effective solutions.

Clinical Value of Growth Differentiation Factor 15 Detection in the Diagnosis of Early Liver Cancer Based on Data Mining.

The incidence of liver cancer is increasing year by year, and how to effectively diagnose early-stage liver cancer and improve the survival rate of liver cancer patients has become one of the current research topics of concern. Aiming at this problem, it is of great significance for the diagnosis of early liver cancer. With the in-depth research on the diagnosis of early-stage liver cancer, the research on growth differentiation factor 15 is gradually carried out, and its performance advantages are of great significance to solve the problem of detection and diagnosis of early-stage liver cancer. This study can improve the accuracy of early diagnosis of liver cancer. The purpose of this paper is to study the application of data mining in the study of clinical value of growth and differentiation factor 15 detection and diagnosis of early liver cancer. In this paper, the data mining algorithm is analyzed, the performance of the algorithm is experimentally analyzed, and the relevant theoretical formulas are used to explain. The results showed that the expression level of GDF-15 was significant in early primary liver cancer (tumor diameter <2.5 cm). Different from normal liver tissue (P < 0.01), there was a significant difference (P < 0.01) compared with adjacent tissue (P < 0.01). Serum GDF-15 can be used as a tumor marker for predicting early stage liver cancer. The high expression of GDF-15 in early stage liver cancer is an independent risk factor affecting the prognosis of liver cancer patients.

Multivariate analysis of serum surface-enhanced Raman spectroscopy of liver cancer patients

Early diagnosis of liver cancer plays a significant role in reducing its high mortality. In this preliminary study, the feasibility of using serum surface-enhanced Raman spectroscopy (SERS) to identify liver cancer was studied. Serum samples were obtained from liver cancer patients and healthy controls. The differences between the SERS spectra of pre-operation and post-operation of liver cancer patients were also analyzed. The general shape and trend of SERS spectra of health control and liver cancer patients were similar. Multivariate analysis, e.g., PLS-SVM, might be useful for the discrimination of serum SERS spectra of pre-operation and post-operation.

An ultrasensitive immunosensor based on cellulose nanofibrils/polydopamine/Cu-Ag nanocomposite for the detection of AFP

In this work, an ultrasensitive immunosensor for amperometric determination of alpha-fetoprotein (AFP) was developed utilizing Ag and Cu nanoparticles on polydopamine (PDA) functionalized cellulose nanofibrils (CNFs) composite (CNFs/PDA/Cu-Ag) as signal amplifier. PDA was first prepared by self-polymerizing of dopamine, and then was adsorbed on CNFs. The obtained CNFs/PDA was applied as substrate to electrolessly deposit Cu-Ag nanoparticles, using NaBH4 as reducing agent. The structure and morphology of the synthesized CNFs/PDA/Cu-Ag nanocomposite were analyzed through Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, X-ray powder diffraction, scanning electron microscopy, particle size analyzer and transmission electron microscopy. The CNFs/PDA/Cu-Ag modified glassy carbon electrode can fix AFP antibody (Ab), and further capture AFP specifically. Electrochemical impedance spectroscopy and cyclic voltammetry were used to characterize the assembly process of immunosensor. The immunoreaction was amplified by electrocatalytical reduction of H2O2 on Cu-Ag nanoparticles, through which AFP was quantitatively detected. The developed sensor exhibits wide linear range of 0.01–100 ng mL−1 (R2 = 0.9963) with low detection limit of 4.27 pg mL−1 (S/N = 3). In addition, it has been used for the detection of AFP in human serum, manifesting its preeminent application prospect in early liver cancer diagnosis.

Establishment and Analysis of a Combined Diagnostic Model of Liver Cancer with Random Forest and Artificial Neural Network

The incidence of liver cancer (hepatocellular carcinoma; HCC) is rising and with poor clinical outcome expected, a more accurate judgment of tumor tissues and adjacent nontumor tissues is necessary. The aim of this study was to construct a diagnostic model based on random forest (RF) and artificial neural network (ANN). It can be used to aid in the identification of diseased tissue such as cancerous tissue, for HCC clinical diagnosis and surgical guidance. GSE36376 and GSE121248 from Gene Expression Omnibus (GEO) were used as training sets in this investigation. R package “limma” and WGCNA were used to filter the training set for statistically significant p &lt; 0.05 differential genes. To better understand the biological function and characteristics, R software was used to perform GO and KEGG enrichment analyses. To pick out and further understand the key genes, we performed PPI analysis and random forest tree analysis. Next, we built the ANN to predict training sets and validation set (GSE84402), and ROC curve was plotted to calculate area under curve (AUC). Then immune cell infiltration indicated difference of immune cell subsets between control and case groups. Finally, the survival analysis of key genes was also carried out based on data in TCGA database. Based on the expression of these 9 genes, we built the artificial neural network (ANN) and the accuracy of the final models was assessed with an ROC curve. The areas under the ROC curve were 0.984 (95% CI 0.972–0.993) in training sets. Its predictive capability was further assessed using the validation set. And the areas under the ROC curve were 0.929 (95% CI 0.786–1.000). In summary, this method effectively classifies hepatocellular carcinoma tissues and the corresponding noncancerous tissues and provides reasonable new ideas for the early diagnosis of liver cancer in the future.

A Novel Fluorescent Probe Strategy Activated by β-Glucuronidase for Assisting Surgical Resection of Liver Cancer.

Liver cancer is a primary malignant tumor with a very high fatality rate, which has seriously threatened human health and life. In normal hepatocellular lesions, β-glucuronidase (GLU) activity in liver cancer tissues is significantly increased. Therefore, GLU has become one of the important biomarkers of primary liver cancer. Here, a series of fluorescent probes (DCDH, DCDCH3, DCDOCH3, and DCDNO2) for early diagnosis of liver cancer and auxiliary surgical resection were successfully synthesized. Since the electron-withdrawing group -NO2 connected to the probe DCDNO2 accelerates the rapid cleavage of the glycosidic bond, DCDNO2 exhibits superior fluorescence properties that are more sensitive and rapid than the other three probes DCDH, DCDCH3, and DCDOCH3 when detecting GLU. DCDNO2 has been well-applied in real-time fluorescent visualization imaging for the detection of GLU activity in liver cancer cells and tumor tissues. In addition, DCDNO2 has also been successfully used in the early diagnosis of liver cancer and real-time imaging to guide the surgical resection of liver cancer tumors. Therefore, DCDNO2 has great potential for development in bioclinical medicine for the early detection and treatment of liver cancer.

CNT biodevices for early liver cancer diagnosis based on biomarkers detection- a promising platform

Regarding the serious threat of liver cancer owing to the concealment and hard detection of liver tumors at an early stage, primary diagnosis becomes quite crucial to guarantee human health. So, in this work platinum-decorated single-walled carbon nanotubes (SWCNTs) were proposed as superior nanodevice for the detection of 1-Octen-3-ol (octenol), decane, and hexanal as liver cancer biomarkers in the exhaled breath of the patients. Herein, density functional theory (DFT) calculations have been utilized to scrutinize the structural and electronic properties of pristine and Pt-decorated SWCNTs. Obtained results showed that the gas molecules were weakly physisorbed on the pristine SWCNT with negligible charge transfer and large interaction distances. Contrariwise, after the decoration of the SWCNT with Pt metal atom, significant charges are transferred, and energy adsorption increased. The results disclosed that the energy adsorption has been enhanced, for example, energy adsorption increased two times for decane and hexanal molecules (−1.06, and −1.07 eV) upon adsorption on Pt-decorated SWCNT. Moreover, substantial charges with amount of 0.238, 0.245, and 0.223 e were transferred from octenol, decane, and hexanal to the surface, respectively. So, investigations revealed that these compounds are strongly chemisorbed on Pt-SWCNT with small interaction distances and along with the short recovery time of 1.7, 83.4, and 123 s at room temperature toward octenol, decane, and hexanal, respectively which make it a compelling nanodevice. Considering the findings, Pt-SWCNT is an excellent substrate for the sense of liver cancer biomarkers with desired recovery time and the results demonstrate its feasibility for potential application in the near future in the field of liver cancer diagnosis.

Proton Magnetic Resonance Spectroscopy at 3.0T in Rabbit With VX2 Liver Cancer: Diagnostic Efficacy and Correlations With Tumor Size.

PurposesThe purpose of this study was to explore the diagnostic performance of Cho peak area (Cho Are), Cho peak amplitude (Cho Amp), and the combined approach (Cho Are_Amp) in detecting rabbit VX2 liver cancer at the early stage via hydrogen-1 proton magnetic resonance spectroscopy (1H-MRS), as well as the correlations between Cho Are, Cho Amp, and tumor parameters like diameter and volume.MethodsConventional magnetic resonance imaging (MRI) and MRS were performed to scan the VX2 liver cancer in rabbit. The tumor diameter was measured on T2-weighted imaging (T2WI), and the tumor volume was accordingly calculated. Cho Are and Cho Amp were obtained from MRS. The diagnostic performance of Cho Are, Cho Amp, and Cho Are_Amp was assessed by a receiver operating characteristic (ROC) curve and area under ROC curve (AUC), whereas specificity and sensitivity were calculated by the maximum Youden’s index. Spearman’s correlation analysis was performed to evaluate the relevance between tumor parameters (diameter, volume) and radiological indexes (Cho Are, Cho Amp).ResultsROC curve analysis showed that Cho Amp, Cho Are, and Cho Are_Amp were effective in diagnosing VX2 liver cancer. The AUC of Cho Amp was 0.883, and the specificity and sensitivity were 0.944 and 0.722, respectively (p < 0.001). The AUC of Cho Are was 0.807, and the specificity and sensitivity were 0.778 and 0.833, respectively (p < 0.05). The AUC of Cho Are_Amp was 0.892, and the specificity and sensitivity were 0.833 and 0.833, respectively (p < 0.001). Cho Are and Cho Amp exhibited a high positive correlation with tumor diameter and tumor volume, among which Cho Amp demonstrated better correlations to tumor diameter and tumor volume (r = 0.956 and 0.946) than that of Cho Are (r = 0.787 and 0.794). A high positive correlation was detected between Cho Are and Cho Amp (r = 0.787), as well as tumor diameter and tumor volume (r = 0.965).ConclusionCho Are_Amp can be used as an effective tool in diagnosing early-stage VX2 liver cancer with satisfied diagnostic accuracy. Cho Are and Cho Amp were positively correlated with tumor volume and tumor diameter. The results of this study provide further evidence that Cho Amp and Cho Are_Amp of MRS could aid in the early diagnosis of liver cancer.

Liver proliferating cell nuclear antigen, BAX/Bcl-2 ratio, collagen, and polysaccharide accumulation as diagnostic tools in experimental hepatocellular carcinoma

Objectives: This experimental study was conducted to look for a sensitive diagnostic panel for early detection of HCC.&#x0D; Methods: Combination of diethyl nitrosamine (DENA, 200 mg/kg, IP, once), two weeks later, CCl4(3ml/kg/week), subcutaneously, for 6 weeks) induced HCC in rats. Sixteen male Wistar rats were divided into 2 groups, control, and HCC.&#x0D; Results: DENA plus CCl4 elevated serum alpha-fetoprotein, liver enzyme activity and depressed superoxide dismutase, reduced glutathione (GSH), elevated malondialdehyde. Also, depressed caspase-3, elevated collagen, polysaccharide accumulation, proliferating cell nuclear antigen in liver tissues, and depressed BAX / Bcl-2 ratio. &#x0D; Conclusion: Decreased BAX/Bcl-2 ratio, elevated collagen deposition, polysaccharide accumulation, nuclear proliferation, and tissue oxidative stress help in the early diagnosis of liver cancer.