|Year : 2020 | Volume
| Issue : 3 | Page : 421-426
Serum Beclin 1 in HCC and correlation with MDA as an oxidant
Zeinab A Mohammed1, Fatma A.K Attia1, Fatma M El-Senosy1, Eman R Younes2, Fadila M Elsayed3
1 Department of Internal Medicine, Al Azhar University, Cairo, Egypt
2 Department of Department of Medical Biochemistry, Medical Division National Research Center, Cairo, Egypt
3 Department of Diagnostic Radiology, Faculty of Medicine for Girls, Al Azhar University, Cairo, Egypt
|Date of Submission||07-Jun-2020|
|Date of Decision||20-Jun-2020|
|Date of Acceptance||23-Jun-2020|
|Date of Web Publication||2-Oct-2020|
MBBCH Zeinab A Mohammed
Department of Internal Medicine, Al Azhar University, Faculty of Medicine (For Girls), AL-Azhar University, Al_Mansoura, Al Dakhlia, 35511
Source of Support: None, Conflict of Interest: None
Background Hepatocellular carcinoma (HCC) is the second most lethal cancer worldwide with persistently increasing mortality in Europe, North/South America, and Africa, in contrast to the decreasing trend in East Asia. Cirrhosis was estimated to cause more than 1.2 million deaths (2% of global deaths) in 2013, an increase of 47% since 1990.
Aim The aim was to assess serum Beclin 1 as a biomarker in HCC regarding its role in pathogenesis and its correlation with serum malondialdehyde (MDA) as an oxidant.
Patients and methods This is a case–control study conducted on 60 participants who were divided into two groups: group 1 included 30 patients with HCC, comprising 26 males and four females, with ages ranging from 55 to 75 years. Group II included 30 healthy participants as a control group, comprising 21 males and nine females, with ages ranging from 25 to 42 years. Routine laboratory investigations were done, and serum Beclin 1 and serum MDA were measured in both groups. Abdominal ultrasonography and triphasic computed tomography were done for the patient group. All patients were recruited from Internal Medicine Department (Al-Mansoura University Hospital) after obtaining oral consent to be participated in the study.
Results There was a highly significant decrease in serum Beclin 1 levels (<0.001) in patients with HCC when compared with the control group. The significant association between Beclin 1 and HCC suggests that low Beclin 1 levels may play an important role in the development of HCC. Moreover, there was a statistically significant increased serum MDA level (>0.001) in patients with HCC as compared with control group. There was a positive correlation between Beclin 1 and BMI in the patient group (r=0.42 and P=0.02), a negative correlation between MDA and red blood cells in the patient group (r=−0.40 and P=0.03), and a positive correlation between MDA and aspartate transaminase in the patient group (r=0.41 and P=0.02).
Conclusion Findings of our study have demonstrated that serum Beclin 1 and MDA levels could be used as possible predictors of pathogenesis of HCC.
Keywords: hepatocellular carcinoma, malondialdehyde, oxidant, serum Beclin 1
|How to cite this article:|
Mohammed ZA, Attia FA, El-Senosy FM, Younes ER, Elsayed FM. Serum Beclin 1 in HCC and correlation with MDA as an oxidant. Sci J Al-Azhar Med Fac Girls 2020;4:421-6
|How to cite this URL:|
Mohammed ZA, Attia FA, El-Senosy FM, Younes ER, Elsayed FM. Serum Beclin 1 in HCC and correlation with MDA as an oxidant. Sci J Al-Azhar Med Fac Girls [serial online] 2020 [cited 2020 Oct 26];4:421-6. Available from: http://www.sjamf.eg.net/text.asp?2020/4/3/421/296951
| Introduction|| |
Hepatocellular carcinoma (HCC) is the second most lethal cancer worldwide with persistently increasing mortality in Europe, North/South America, and Africa in contrast to the decreasing trend in East Asia. Cirrhosis was estimated to cause more than 1.2 million deaths (2% of global deaths) in 2013, an increase of 47% since 1990. Cirrhosis and HCC are the major life-limiting consequences of progressive fibrotic liver diseases, mainly caused by hepatitis B virus (HBV), hepatitis C virus, alcohol abuse, and nonalcoholic fatty liver disease .
Beclin 1 is a 60 kDa coiled-coil protein. The function of Beclin 1 in autophagy was first suspected owing to its amino acid sequence identity in the yeast autophagy protein (Atg6). Beclin 1 was found to restore autophagic activity in Atg6-disrupted yeast, becoming one of the first identified mammalian genes to regulate autophagy.
Beclin 1-depleted cells cannot induce autophagosome formation. Beclin 1 in humans is encoded by the BECN1 gene, which is located on chromosome 17 .
Malondialdehyde (MDA) is the organic compound with the nominal formula CH2(CHO)2. A colorless liquid, MDA is a highly reactive compound that occurs as the enol. MDA is the main product of membrane lipid peroxidation; it can also be generated during prostaglandin biosynthesis in cells . This MDA-induced DNA alteration may contribute significantly to many cancers, including HCC .
| Aim|| |
The aim was to assess serum Beclin 1 as a biomarker in HCC regarding its role in pathogenesis and its correlation with serum MDA as an oxidant.
| Patients and methods|| |
This study includes 60 Egyptian patients with HCC, comprising 26 males and four females, with age ranging from 55 to 75 years (group 1). Also approval of research ethical committee of faculty of medicine for girls-AlAzhar university was obtained. Moreover, 30 apparently healthy volunteers were included in the study as a control group (group II), comprising 21 males and nine females, with age ranging from 25 to 42 years. All patients were recruited from Internal Medicine Department (Al-Mansoura University Hospital) after obtaining oral consent to be participated in the study.
Patients with chronic kidney disease, patients with uncontrolled diabetes or autoimmune diseases, patients with heart failure, patients with hypertension, obese patients, and patients with other malignancy all over the body were excluded from the study.
All participants in this study were subjected to the following:
Full medical history and complete physical examination.
Measurement of BMI: weight divided by the square of the height (kg/m2) laboratory investigations:
- Complete blood picture, liver function tests [serum albumin, bilirubin, alanine transaminases and aspartate transaminase (AST)], and kidney function tests including serum urea and creatinine.
- Specific investigations:
- Beclin 1 was estimated by enzyme-linked immunosorbent assay, based on the sandwich technique, according to Sino Geneclon Biotech Co. Ltd (Hangzhou, China) for research, catalog No: SG 00353.
- Lipid peroxidation in serum (MDA) was assayed by measuring the level of MDA using the method of Ruiz-Larrea and colleagues. In this assay, thiobarbituric acid (TBA) reactive substances (primarily MDA) react with TBA to form TBA-MDA adduct, which can be measured colorimetrically at 532 nm.
- Radiologic investigations:
- Abdominal ultrasonography with special emphasis on the following:
- Liver size, echo pattern and presence of focal lesion, splenic size and echo pattern, portal vein diameters, and presence or absence of ascites.
- Triphasic computed tomography abdomen.
Data were collected, revised, coded, and entered to the Statistical Package for the Social Sciences (IBM SPSS, Missouri, USA) version 15. Quantitative data were presented as mean±SD. Qualitative data were expressed as frequency and percentage. Probability (P value) was set as follows: P value less than 0.05 was considered significant, P value less than 0.001 was considered as highly significant, and P value more than 0.05 was considered insignificant.
| Results|| |
Demographic data of the studied groups
The age of group I (30 patients with HCC) ranged from 55 to 75 years, with mean±SD of 63.97±3.44 years, and BMI ranged from 19 to 25, with mean±SD of 25.31±1.96. The age of group II (30 healthy volunteers as a control group) ranged from 25 to 42 years, with mean±SD of 35.27±4.35 years, and BMI ranged from 18 to 25, with mean±SD of 25.94±4.59.
Comparison between group 1 and group II
Regarding complete blood count there was a significant decrease in hemoglobin and platelet in group I (8.84±1.82 g/dl and 107.73±19.67×103, respectively) when compared with group II (13.52±1.26 g/dl and 300.33±88.02×103, respectively) ([Table 1]). However, there was a nonsignificant increase in white blood cells and RBCs in group I when compared with group II. ([Table 1]).
|Table 1 Comparison between group I and group II with respect to the studied parameters|
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There was a significant increase in total serum bilirubin and direct bilirubin in group I (4.23±1.56and 1.99±081 mg/dl, respectively) when compared with group II (1±.0 and 016±0.07 mg/dl, respectively). However, there was a significant decrease of serum albumin in group I (3.20±0.66 g/dl) when compared with group II (4.57±0.60 g/dl) ([Table 1]).
There was a highly significant increase of alanine transaminase and AST in group I (48.93±12.09 and 59.50±17.15 μ/l, respectively) when compared with group II (25.57±6.08 and 23.90±5.86 μ/l, respectively) ([Table 1]).
There was a highly significant increase of PT, PC, and INR in group I (21.83±3.96 second, 55.77±10.84%, and 2.42±0.60, respectively) when compared with group II (12.63±1.47 second, 99.67±0.99%, and 1.01±0.03, respectively) ([Table 1]).
Regarding renal function tests
There was a significant increase of serum creatinine and urea in group 1 (1.06±033 and 32.80±7.69 mg/dl when compared with group 2 (0.72±017 and 28.70±4.64 mg/dl, respectively) ([Table 1]).
However, there was a significant decrease of serum Beclin 1 in group I (120.33±28.60 pg/ml) when compared with group II (141.45±45.19 pg/ml) (P=0.03), whereas there was a significant increase of serum MDA in group I (6.32±3.58 nmol/ml) when compared with group II (2.91±0.42 nmol/ml) ([Table 1]).
Correlation of Beclin 1 with the studied parameters in group 1 showed that there was a significant positive correlation between Beclin 1 and BMI ([Figure 1]), whereas there was a nonsignificant negative correlation between Beclin 1 and each of red blood cells (RBCs), white blood cells, and hemoglobin level. In addition, serum Beclin 1 was nonsignificantly positively correlated with serum albumin and MDA ([Table 2]).
|Table 2 Correlation study between Beclin 1 and MDA vs other studied parameters in the patient group|
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Correlation of MDA with the studied parameters in group 1showed that there was a significant negative correlation between MDA and RBCs in the patient group ([Figure 2]), whereas MDA is significantly positively correlated with AST in the patient group ([Table 2]).
| Discussion|| |
This study was carried out to estimate serum Beclin 1 as a biomarker in HCC regarding its role in pathogenesis and its correlation with serum MDA as an oxidant.
This work was conducted on 60 individuals divided into two groups:
- Group I: 30 patients with patients with HCC, as the case group.
- Group II: 30 healthy controls, as the control group.
In our study, we observed a significant decrease in serum Beclin 1 levels in patients with HCC compared with the control group. The significant association between Beclin 1 and HCC suggests that low Beclin 1 levels may play an important role in the development of HCC.
In support of our results, Zhiqiang Qin et al. , studied 1086 patients with HCC and showed that decreased Beclin 1 expression may signify the poor prognosis of HCC. Hence, activation of autophagy may improve the prognosis of patients with HCC.
Moreover, this is in agreement with Jia et al. , who demonstrated that BECN1 deletion is observed in 40–75% human cancers. Interestingly, a heterozygous deletion of atg6/becn1 in mice resulted in increased tumorigenesis in multiple tissues, including the liver. Moreover, BECN1 deletion accelerated HBV-related HCCs.
Moreover, our results are in agreement with Macek Jilkova et al. , who studied animal models of HCC and demonstrated that genetic models have shown that inhibition of autophagy genes (as Beclin 1 gene) favors the development of liver tumors. Not only do mice heterozygous for Beclin 1 show a high frequency of spontaneous HCC, they also display accelerated development of HBV-induced small-cell dysplasia. Accordingly, the expression of Beclin 1 is lower in HCC cells than in adjacent non-tumor cells. Inhibition of autophagy leads to induction of tumorigenesis.
The results are also in agreement with Shravage et al. , who demonstrated that most homozygous Atg6 (Beclin 1) null mutant animals die at the end of larval development. Atg6 is a core component of the Vps34 complex, and studies in yeast and mammalian systems have identified Vps34 as an essential protein regulating a wide variety of vesicular trafficking events, including autophagy and endocytosis. Beclin 1 plays an important role in cancer, and most studies attribute this function to a defect in autophagy.
These results are in agreement with Akkoç and Gözüaçık , who showed that several autophagy-deficient animal models have been shown to experience hepatic steatosis and demonstrated that autophagy is involved in lipid and glycogen metabolism. Evidently, these and other abnormalities of liver function and pathologies are also closely related to HCC development.
In support of our results, Marinković et al.  demonstrated that the tumor-suppressing role of autophagy in HCC has been confirmed on several HCC models. One of the first evidence supporting the tumor suppressor role of autophagy in the cancer formation comes from Beclin 1 knockout mice. Beclin 1 deletion reduces autophagy activity, and such mice were more likely to develop cancer, including HCC.
We are also in agreement with Sheng-Hui Lan et al. , who showed that there are many accumulating evidence suggesting that autophagy is involved in tumor suppression. Autophagy-deficient mice develop multiple liver tumors and promote fibrosarcomas induced by chemical carcinogens. Low autophagic activity was detected in HBV-associated HCC specimens and HBx-transgenic mice.
In support of our results, Dong-Mei Qiu et al. , showed that the expression of the autophagic gene Beclin 1 and its autophagic activities are suppressed in some HCC tissues. Autophagy defects may be associated with tumor progression and poor prognosis of HCC. The expression of Beclin 1 may be a valuable marker to estimate HCC progression.
In the present study, we observed significant increased serum MDA levels in patients with HCC as compared with control group. The significant association between MDA and HCC suggests that high MDA levels may play an important role in the development of HCC.
This is in agreement with Matlab and Jasim , who recorded high concentrations of MDA in patients with HCC and those of liver transplantation patients in comparison with non-tumoral liver diseases.
It was also in agreement with Unsal and Belge-Kurutaş , who showed that ROS contributes to the initiation and progression of HCC, and MDA appears to be the most mutagenic product of lipid peroxidation. MDA is one of the most popular and reliable markers that determine oxidative stress in research.
In support of our results, Cheng et al.  demonstrated that carcinogenesis may progress or be enhanced if an imbalance between oxidative stress and antioxidant defense capacities occurs. Our patients had increased oxidative stress (MDA levels) before they underwent HCC resection. However, the expression of oxidative stress and antioxidant markers was reversed after HCC resection. This supports that increased oxidative stress is an important pathogenic mechanism of HCC.
We are in agreement with Liu et al. , who showed that after one month of the surgery, patients with HCC exhibited significantly lower oxidative stress and higher antioxidant enzymes activities. The level of oxidative stress (MDA) was significantly reduced by 15%, and the antioxidant activities were significantly increased.
In support of our results, Gaschler and Stockwell , showed that when oxidant compounds target lipids, they can initiate lipid peroxidation process, which produces multiple breakdown molecules, such as MDA. Among several substrates, proteins and DNA are particularly susceptible to modification caused by these aldehydes.
Our study is also in agreement with Hassan et al. , who showed that there is a significant relationship between HCC and free radical-mediated oxidative stress demonstrated by increased levels of MDA as well as decreased levels of antioxidant parameters in the examined organs of rats. Thus, blocking the oxidative stress pathway may be of therapeutic value in treatment of liver injury.
In support of our results, Saieva et al. , showed that MDA is a mutagenic and carcinogenic compound, being able to react with DNA, forming specific DNA adducts such as M1dG. These adducts can be considered a suitable biomarker of DNA damage, induced endogenously by lipid peroxidation and also exogenously by direct oxidation of DNA from ROS by the pathway of the oxidative stress.
Our study is in agreement with Lin et al. , who found that plasma lipid oxidation level (MDA) in patients with HCC was significantly greater than healthy participants.
In our study, serum Beclin 1 showed a significant positive correlation with BMI,
Our result was in agreement with Namkoong et al. , who showed that autophagy is one of the major degradative mechanisms that can eliminate excessive nutrients, toxic protein aggregates, damaged organelles, and invading microorganisms. The results were also in agreement with Madrigal-Matute and Cuervo , who showed that although nutritional abundance can generally suppress autophagy, oxidative stress insults associated with obesity can stimulate autophagy as a stress defense mechanism.
Our results disagree with Tripathi et al. , who showed that obesity suppresses autophagy (Beclin 1) and allows accumulation of undesired proteins. If not removed properly, these accumulations induce pathological conditions.
In our study, serum MDA shows a significant negative correlation with RBCs.
This is in agreement with Barodka et al. , who showed that red blood cell oxidative stress impairs oxygen delivery and induces red blood cell aging. Oxidative stress (MDA) contributes to the uptake of RBCs by macrophages, which play a major role in the removal of RBCs from circulation. The contribution of oxidative stress to the removal of RBCs by macrophages involves caspase-3 activation, which requires oxidative stress. So, RBC oxidative stress plays a significant role in inducing RBC aging and decreasing RBCs.
In our study, serum MDA showed a significant positive correlation with liver enzyme AST.
In support to our results Contreras and Hernández 2016  who demonstrated that there is an increased production of ROS, leading to hepatic lipid peroxidation and oxidative stress which activate Kupffer cells and initiate the inflammatory response and development of fibrosis. This evidence strongly suggests the participation of oxidant stress in acute liver damage, probably inducing the progression of liver injury to chronic liver damage.
In agreement with our results, Nazeri et al.  showed that AST exists in a variety of tissues such as liver, heart, muscle, kidney, and brain. When these tissues are damaged by oxidative stress, this enzyme is secreted and increased in the blood.
| Conclusion|| |
Our study has demonstrated that serum Beclin 1 and MDA levels could be used as possible predictors of pathogenesis of HCC.
Serum Beclin 1 and serum MDA could be used as a marker for prediction of HCC.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Fujiwara N, Friedman SL, Goossens N, Hoshida Y. Risk factors and prevention of hepatocellular carcinoma in the era of precision medicine. J Hepatol 2018; 68:526–549.
Hayat M. Overview of autophagy. In: Autophagy: Cancer, Other Pathologies, Inflammation, Immunity, Infection, and Aging. USK: Academic Press; 2017. 12:1–122.
Matlab N, Jasim R. Assessment of the cellular balance for production of oxidants-antioxidants in serum samples of patients with advanced stages of cancer tumors. J Appl Chem 2017; 6:433–447.
Martinez-Useros J, Garcia-Foncillas J. Obesity and colorectal cancer: molecular features of adipose tissue. J Transl Med 2016; 14:21.
Zhiqiang Qin Z, Yu X, Lin M, Wu J, Ma S, Wang N. Prognostic and clinicopathological value of Beclin 1 expression in hepatocellular carcinoma: a meta-analysis. World J Surg Oncol 2018; 16:170.
Jia J, Zhang HB, Shi Q, Yang C, Ma JB, Jin B, Guo P. KLF5 downregulation desensitizes castration-resistant prostate cancer cells to docetaxel by increasing BECN1 expression and inducing cell autophagy. Theranostics 2019; 9:5464.
Macek Jilkova Z, Kurma K, Decaens T. Animal models of hepatocellular carcinoma: the role of immune system and tumor microenvironment. Cancers 2019; 11:1487.
Shravage BV, Hill JH, Powers CM, Wu L, Baehrecke EH. Atg6 is required for multiple vesicle trafficking pathways and hematopoiesis in Drosophila. Development 2013; 140:1321–1329.
Akkoç Y, Gözüaçık D. Autophagy and liver cancer. Turk J Gastroenterol 2018; 29:270.
Marinković M, Buljubašić M, Novak I, Šprung M. Autophagy modulation in cancer: current knowledge on action and therapy. Oxid Med Cell Longev 2018; 2018:443–447.
Lan S-H, Wu S-Y, Zuchini R, Lin XZ, Su IJ, Tsai TF, Liu HS. Autophagy suppresses tumorigenesis by degrading oncogenic miR-224 in HCC. (No. 97-EC-17-A-31-F1-0695). 2017.
Qiu D-M, Wang G-L, Chen L, Xu YY, He S, Cao XL, Qun E. The expression of beclin-1, an autophagic gene, in hepatocellular carcinoma associated with clinical pathological and prognostic significance. BMC Cancer 2014; 14:327.
Unsal V, Belge-Kurutaş E. Experimental hepatic carcinogenesis: oxidative stress and natural antioxidants. Open Access Maced J Med Sci 2017; 5:686.
Cheng SB, Liu HT, Chen SY, Lin PT, Lai CY, Huang YC. Changes of oxidative stress, glutathione, and its dependent antioxidant enzyme activities in patients with hepatocellular carcinoma before and after tumor resection. PLoS ONE 2017; 12:345.
Liu HT, Cheng SB, Huang YC, Huang YT, Lin PT. Coenzyme Q10 and oxidative stress: Inflammation status in hepatocellular carcinoma patients after surgery. Nutrients 2017; 9:29.
Gaschler MM, Stockwell BR. Lipid peroxidation in cell death. Biochem Biophys Res Commun 2017; 482:419-425.
Hassan HA, Ghareb N, Azhari G. Antioxidant activity and free radical-scavenging of cape gooseberry (Physalis peruviana
L.) in hepatocellular carcinoma rats model. Hepatoma Res 2017; 3:27–33.
Saieva C, Peluso M, Palli D et al.
Dietary and lifestyle determinants of malondialdehyde DNA adducts in a representative sample of the Florence City population. Mutagenesis 2016; 31:475–480.
Lin C, Yin M. B vitamins deficiency and decreased anti-oxidative state in patients with liver cancer. Eur J Nutr 2007; 46:293–299.
Namkoong S, Ho A, Woo Y et al.
Systematic characterization of stress-induced RNA granulation. Mol Cell 2018; 70:175–187.
Madrigal-Matute J, Cuervo A. Regulation of liver metabolism by autophagy. Gastroenterology 2016; 150:328–339.
Tripathi S, Srivastava S, Tripathi Y. Obesity and its complications: role of autophagy. Int J Pharm Sci Res 2018; 9:3100–3113.
Barodka VM, Nagababu V, Mohanty JG et al.
New insights provided by a comparison of impaired deformability with erythrocyte oxidative stress for sickle cell disease. Blood Cells Mol Dis 2014; 52:230–235.
Contreras-Zentella M, Hernández-Muñoz R. Is liver enzyme release really associated with cell necrosis induced by oxidant stress? Oxid Med Cell Longev 2016; 2016:241.
Nazeri S, Farhangi M, Modarres S. The effect of different dietary inclusion levels of rutin (a flavonoid) on some liver enzyme activities and oxidative stress indices in rainbow trout, Oncorhynchus mykiss (Walbaum) exposed to Oxytetracycline. Aquac Res 2017; 48:4356–4362.
[Figure 1], [Figure 2]
[Table 1], [Table 2]