Expression of c-erb-B2 gene in bladder cancer of Egyptian patients and its correlation with p53 and bcl-2

doi:10.1016/j.biopha.2015.10.021

Biomedicine & Pharmacotherapy

Volume 76, December 2015, Pages 73-81

https://doi.org/10.1016/j.biopha.2015.10.021 Get rights and content

Abstract

Urinary bladder cancer is the 9th most common type of cancer and the 13th most common cause of death worldwide. C-erbB-4 is a class of oncogenes plays a role in cancer development. The present work was performed to assess C-erbB-4 oncogene amplification by PCR technology and its correlation with p53 and bcl-2. This study included 50 male patients (10 controls and 40 urinary bladder cancer patients). The bladder cancer patients include 20 specimens diagnosed as transitional cell carcinoma (TCC) and 20 specimens diagnosed as squamous cell carcinoma (SCC). The results revealed that 7 (35%) of both TCC and SCC showed c-erb-B2 gene amplification. 12 (60%) of TCC and 6 (30%) of SCC showed positive expression of p53. 11 (55%) of TCC and 6 (30%) of SCC showed positive Bcl-2 expression. A direct statistically significant association was detected between c-erb-B2 expression and Bcl-2 and p53 expression in TCC and SCC specimens. Seven (35%) of TCC showed c-erb-B2 gene amplification and expression of both p53 and Bcl-2. Five (25%) of the examined SCC specimens showed c-erb-B2 gene amplification and positive expression for both p53 and Bcl-2. The results indicated that a direct statistically significant association was detected in TCC group between amplification of c-erb-B2 gene by PCR and expressionof p53 and Bcl-2 by immunohistochemistry.

Introduction

Urinary bladder cancer (UBC) is the 9th most common type of cancer and the 13th most common cause of death worldwide. The highest incidence rates of UBC were found in Europe, United States, and Egypt [1]. The age standardized rates of UBC are 16.9/100,000 (in more developed countries) and 5.3/100,000 (in less developed countries) compared to 21.1/100,000 in Egypt. In Egypt, UBC is one of the most common malignancies in males and ranked 2nd among them [2]. Urinary bladder cancer rates in men are three to four times than those in females [3]. The two main histological types of UBC are urothelial or transitional cell carcinoma (TCC) and squamous cell carcinoma (SCC). The other varieties represent small percentage i.e. less than 5–10% [4].

The tumor suppressor protein p53 is a key regulator of the cellular response to genotoxic damage, and thus plays a pivotal role in preventing cancer formation. Once DNA damage has been incurred, p53 can elicit several different responses to either correct the error(s) or destroy the damaged cell. First, p53 can induce the first growth phase cell cycle arrest, which stops the cell from dividing and allows time to repair the damage before the DNA is replicated. Second, p53 can activate DNA repair proteins to derive the repair of damaged DNA. Third, as a last resort, p53 can induce damaged cells to undergo apoptosis, thereby eliminating damaged and potentially dangerous cells at risk for neoplastic transformation. Due to its critical importance in maintaining genetic stability, p53 has been called the “gatekeeper” or “guardian” of genome [5]. The bcl-2 family proteins are the most important regulators of the mitochondrial pathway of apoptosis [6]. bcl-2 family proteins are divided into three subfamilies, bcl-2, bcl-2-associated X protein (Bax)/bcl-2 homologous antagonist killer (Bak) and BH3-only subfamilies [7]. bcl-2 subfamily proteins have BH1, 2 and 3 and promote cell survival (anti-apoptotic bcl-2 family). Bax/Bak subfamily proteins have BH1, 2 and 3 and are pro-apoptotic. BH3-only subfamily proteins are also pro-apoptotic and function as inhibitors of anti-apoptotic bcl-2 family proteins or activators of Bax/Bak.

The human epidermal growth factor (HER2, also known HER2/Neu or c-erb-B2) gene is located on the chromosome 17q. It is a proto-oncogene, a normal gene with potential to become an oncogene as a result of molecular alterations such as mutation, amplification or overexpression of its protein product [8]. It encodes a transmembrane growth factor receptor with tyrosine kinase activity that is a member of the epidermal growth factor receptor family [9]. The HER2/neu-encoded protein molecule occupies a critical position in the biochemical pathways responsible for the transduction of mitogenic signals from a variety of growth factor receptors. In addition to its role in regulating normal cellular proliferation, overexpression of the HER2/neu gene appears to play a role in neoplastic cell growth [10]. C-erb-B2 amplification and/or overexpression was detected in a variety of human cancers including bladder cancer [11], [12], [13]. The present work was performed to assess C-erbB-4 oncogene amplification by PCR technology in bladder cancer and its correlation with p53 and bcl-2.

Section snippets

Patient groups

This study included 40 urinary bladder cancer patients obtained from Pathology Department, National Cancer Institute, Cairo University. The study met the criteria of the Ethics committee of the institution. The patients were divided into two groups:

Clinical data

Clinical data related to the selected cases were obtained from the patients’ medical records

Clinicopathological data

The total number of specimens used in the present study was 40 specimens, 20 transitional cell carcinoma and 20 squamous cell carcinoma. The mean ages of TCC and SCC were 46.4 ± 5.5 and 45.4 ± 6.6 years, respectively. The mean tumor sizes of both TCC and SCC were 2.35 ± 0.8 and 2.21 ± 0.5 cm, respectively. In TCC group (20 specimens), 16 specimens (80%) were in grade 2 while only 4 specimens (20%) were in grade 3. Concerning the SCC group (20 specimens), 15 specimens (75%) were in grade 2 while only 5

Discussion

Urinary bladder cancer is the most common malignancy of the urinary tract [15]. In Egypt based on the results of National Population-Based Registry Program 2008–2011, the UBC is the 3rd most frequent cancer in both sexes. It ranked 2nd in males after liver cancer and represented 10.71% [2].

In the present study, no significant difference was recorded between the ages of TCC and SCC groups. Zaghloul et al. [4] reported an increase of age with time in TCC cases from 54.2 ± 9.5 in period 1988–1993 to

References (44)

S. Chavan et al.
International variations in bladder cancer incidence and mortality
Eur. Urol.
(2014)
N.D. Smith et al.
The p53 tumor suppressor gene and nuclear protein: basic science review and relevance in the management of bladder cancer
J. Urol.
(2003)
C.C. Guo et al.
Squamous cell carcinoma of the urinary bladder: a clinicopathologic and immunohistochemical study of 16 cases
Hum. Pathol.
(2009)
E.J. Kirsh et al.
Expression of bcl-2 and bcl-X in bladder cancer
J. Urol.
(1998)
L. Nakopoulou et al.
The prevalence of bcl-2, p53, and Ki-67 immunoreactivity in transitional cell bladder carcinomas and their clinicopathologic correlates
Hum. Pathol.
(1998)
S. Charfi et al.
Immunohistochemical expression of HER2 in urothelial bladder carcinoma and its correlation with p53 and p63 expression
J. Microsc. Ultrastruct.
(2013)
A.S. Ibrahim et al.
Cancer incidence in Egypt: results of the national population-based cancer registry program
J Cancer Epidemiol.
(2014)
H. Fajkovic et al.
Impact of gender on bladder cancer incidence, staging, and prognosis
World J. Urol.
(2011)
M.S. Zaghloul et al.
Time-trend in epidemiological and pathological features of schistosoma-associated bladder cancer
J. Egypt. Natl. Cancer Inst.
(2008)
J.G. Teodoro et al.
P53 and angiogenesis

W.X. Ding et al.

The Bcl-2 family proteins

J. Ninomiya-Tsuji

Mitochondrial dysfunction

K.A. Cadoo et al.

Advances in molecular and clinical subtyping of breast cancer and their implications for therapy

G.P. Kalemkerian et al.

Present concepts in the molecular biology of lung cancer

G. Atış et al.

Determination of HER2/NEU gene amplification and protein overexpression in bladder transitional cell carcinoma

Adv. Mol. Biol.

(2007)

U. Lönn et al.

Prognostic value of amplification of c-erb-B2 in bladder carcinoma

Clin. Cancer Res.

(1995)

J.N. Wiegand et al.

HER2 overexpression and amplification is presented in a subset of ovarian mucinous carcinomas and can be targeted with trastuzumab therapy

BMC Cancer

(2009)

A. Shawky et al.

Her-2/Neu overexpression in invasive bladder carcinoma among a cohort of Egyptian patients

World J. Nephrol. Urol.

(2013)

U. Lönn et al.

Prognostic value of erb-B2 and myc amplification in breast cancer imprints

Cancer

(1995)

A. Anastasiadis et al.

Best practice in the treatment of nonmuscle invasive bladder cancer

Ther. Adv. Urol.

(2012)

D.M. El-Rehim et al.

Expression of extracellular matrix metalloproteinase inducer and fascin in urinary bladder cancer: correlation with clinicopathological characteristics

Mol. Clin. Oncol.

(2013)

N. Lagwinski et al.

Squamous cell carcinoma of the bladder: a clinicopathologic analysis of 45 cases

Am. J. Surg. Pathol.

(2007)

Cited by (10)

Optimal first-line treatment for platinum-eligible metastatic urothelial carcinoma: Comparison of chemo-immunotherapy, immunotherapy, and chemotherapy— A systematic review and meta-analysis
2022, Clinical Immunology
Citation Excerpt :
Urothelial carcinoma (UC) is the most common malignancy involving the genitourinary system and the 9th most common malignancy worldwide [1].
The role of first-line of immunotherapy in metastatic urothelial carcinoma (mUC) remains unclear. This meta-analysis aimed to explore an optimal first-line treatment strategy for mUC patients.
We carried out a meta-analysis between chemo-immunotherapy, immunotherapy, and chemotherapy in mUC based on randomized trials. The outcomes included overall survival (OS), progression-free survival (PFS), objective response rate (ORR), disease control rate (DCR), and treatment-related adverse events (AEs). A fixed-effect or random-effects model was adopted depending on between-study heterogeneity.
Three trials involving 3238 patients were included. PD-1/PD-L1 inhibitor plus platinum-based chemotherapy was associated with the improvements of OS (HR, 0.85; 95% CI 0.75–0.99), PFS (HR, 0.80; 95% CI 0.71–0.90) and ORR (OR, 1.32; 95% CI 1.07–1.63) when compared with platinum-based chemotherapy, but not with better DCR (OR, 1.07; 95% CI 0.78–1.46). PD-1/PD-L1 inhibitor alone was associated with worse ORR (OR, 0.38; 95% CI 0.17–0.87) and DCR (OR, 0.20; 95% CI 0.16–0.25) when compared with platinum-based chemotherapy while it did not statistically reduce the risk of mortality (HR 0.97 for entire cohort; 0.90 for PD-L1 high cohort). In safety analyses, the incidence of adverse events (AEs) between regimens showed no difference, but the frequency of AEs of grade 3 or severity was higher in chemo-immunotherapy compared to chemotherapy.
Compared with platinum-based chemotherapy, chemo-immunotherapy is associated with significantly improved PFS, OS, and ORR in the first-line therapy for mUC at the expanse of increased toxicity.
Zinc antagonizes common carp (Cyprinus carpio) intestinal arsenic poisoning through PI3K/AKT/mTOR signaling cascade and MAPK pathway
2021, Aquatic Toxicology
Citation Excerpt :
TUNEL showed higher fluorescence intensity in As group, and apoptotic bodies and abnormal mitochondria were observed under the electron microscope, preliminary suggesting of the occurrence of apoptosis. And our experiments indicate that p53 was increased significantly under As exposure, which may due to the induction of ROS and the signal sent by DNA damage (Sakr et al., 2015). p53 up-regulated modulator of apoptosis (PUMA), as the downstream of p53, it interacted with Bcl-2 through the BH3 domain, and then release the Bax to mitochondrial membrane.
Arsenic (As) pollution is a serious and longstanding problem, which has obvious threaten to aquatic organisms. The study aimed to explore the mitigation effect of natural antioxidant zinc (Zn) on As toxicity in the foregut and midgut of common carp (Cyprinus carpio L.), and in-depth disclose related signal cascade. Carps were treated with Zn²⁺ (1 mg/L) and/or As³⁺ (2.83 mg/L) for a period of 30 days. Under As exposure, the foregut and midgut showed obvious burst of reactive oxygen species (ROS) and breakdown of antioxidant system. What followed is the activation of the endogenous and exogenous apoptotic pathways, and the rise of autophagy level prompted by the increase in LC3 II and the down-regulation of p62. Mitochondrial swelling, cristae fragmentation and autophagosomes were observed under the electron microscope, which also means the occurrence of apoptosis and autophagy. In addition, As induced the activation of p38 mitogen-activated protein kinase (MAPK), c-Jun N-terminal kinase (JNK) and the inhibition of extracellular signal-related kinase (ERK) in MAPK signaling, and up-regulated the level of autophagy through the inhibition of the phosphatidylinositol 3 kinase (PI3K)/AKT/ mammalian target of rapamycin (mTOR) signaling cascade. However, Zn supplementation has clearly reversed the above phenomenon, and it basically has no effect on foregut and midgut. In conclusion, this study shows that Zn can alleviate the damage caused by subchronic As exposure, which provides a reference for the use of Zn preparations in aquaculture.
The involvement of the mitochondrial pathway in manganese-induced apoptosis of chicken splenic lymphocytes
2016, Chemosphere
Citation Excerpt :
Excess ROS can directly induce DNA damage. DNA damage activates p53 (Sakr et al., 2015). Activated p53 stimulates Bax, leads to mitochondrial dysfunction, and finally results in apoptosis in mice livers (Xu et al., 2008).
The purpose of this study was to investigate the effect of excess manganese (Mn)-induced cytotoxicity on apoptosis in chicken splenic lymphocytes. Chicken splenic lymphocytes were cultured in medium in the absence and presence of manganese (II) chloride (MnCl₂) (2 × 10⁻⁴, 4 × 10⁻⁴, 6 × 10⁻⁴, 8 × 10⁻⁴, 10 × 10⁻⁴, and 12 × 10⁻⁴ mM), in N-acetyl-l-cysteine (NAC) (1 mM), and the combination of MnCl₂ and NAC for 12, 24, 36, and 48 h. Tests were performed on morphologic observation, reactive oxygen species (ROS) and malondialdehyde (MDA) content, manganese superoxide dismutase (Mn-SOD) and glutathione peroxidase (GSH-Px) activities, B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), p53, and calmodulin (CaM) messenger RNA (mRNA) expression, Caspase-3 mRNA and protein expression, intracellular free Ca²⁺ ([Ca²⁺]i), and mitochondrial transmembrane potential (ΔΨm). Our research indicated that excess Mn induced ROS and MDA content, inhibited Mn-SOD and GSH-Px activities, induced Bax and p53 mRNA expression, inhibited Bcl-2 and CaM mRNA expression, induced Caspase-3 mRNA and protein expression, upregulated [Ca²⁺]i, inhibited ΔΨm, and induced apoptosis in a dose effect. NAC relieved excess Mn-caused the changes of all above factors. Mn-induced oxidative injuries were alleviated by treatment with NAC, an ROS scavenger. The above results demonstrated that excess Mn caused oxidative stress and apoptosis via mitochondrial pathway in chicken splenic lymphocytes.
Long noncoding RNA SNHG1 predicts a poor prognosis and promotes hepatocellular carcinoma tumorigenesis
2016, Biomedicine and Pharmacotherapy
Citation Excerpt :
Mechanistically, we found that SNHG1 inhibits p53 expression. p53 is a well known tumor inhibitor, which plays pivotal roles in tumorigenesis and development of many cancers [26,27]. p53 inhibits cell cycle progression and induces cell apoptosis through directly transcriptionally activate or repress target genes expression, such as apoptosis related target genes BAX and FAS, and cell cycle related target gene CDKN1A [28,29].
Hepatocellular carcinoma (HCC) is the main cause of cancer mortality worldwide. Its poor prognosis is mainly ascribed to high recurrence rate. Identifying novel prognostic biomarkers and therapeutic targets would be vital for HCC management. Long noncoding RNA (lncRNA) is a class of RNA with various roles in tumorigenesis. The aim of this study was to investigate the clinical significance and functions of lncRNA-small nucleolar RNA host gene 1 (SNHG1) in HCC. In this study, we found SNHG1 was upregulated in HCC tissues in comparison with adjacent liver tissues in both publicly available microarray data and our own cohort. High SNHG1 expression was correlated with large tumor size, poor differentiation, and aggressive BCLC stage. Kaplan-Meier survival analysis demonstrated that high SNHG1 expression predicts poor prognosis of HCC patients. Gain-of-function and loss-of function experiments showed that SNHG1 promotes HCC cells proliferation, cell cycle progression, and inhibits HCC cells apoptosis. Further experiments revealed that SNHG1 promotes HCC cells proliferation through inhibiting p53 and p53-target genes expression. Collectively, our results demonstrated the clinical prognostic significance and roles of SNHG1 in HCC, and suggested that SNHG1 may be considered as a prognostic biomarker and therapeutic target for HCC.
Perspectives on the future of urothelial carcinoma therapy: chemotherapy and beyond
2023, Expert Opinion on Pharmacotherapy
Effect of long-term manganese exposure on oxidative stress, liver damage and apoptosis in grouper Epinephelus moara ♀ × Epinephelus lanceolatus ♂
2022, Frontiers in Marine Science

View all citing articles on Scopus

View full text

Original articleExpression of c-erb-B2 gene in bladder cancer of Egyptian patients and its correlation with p53 and bcl-2