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Introduction 

Lung cancer is one of the important causes of mortality 
worldwide (Park et al., 2015; Masuda et al., 2015). Non-
small cell lung cancer is the most common subtype, 
accounting for approximately 80-85% of all lung cancers 
(Park et al., 2015; Masuda et al., 2015). Most patients 
with non-small cell lung cancer are diagnosed at the 
advanced stages (stages IIIb and IV), with only 16-30% 
diagnosed in the early stages of the disease (Park et al., 
2015; Masuda et al., 2015; Polanski et al., 2016). Patients 
diagnosed with advanced non-small cell lung cancer 
generally have a poor prognosis, with a median survi-
val of 8-10 months and 2 and 5-year survival rates of 
approximately 20% and 15%, respectively (Park et al., 
2015; Masuda et al., 2015; Polanski et al., 2016; Ali et al., 
2013).  

Poly (ADP-ribose) polymerase-1 (PARP-1) is also 

known as adenosine diphosphate-ribosyltransferases 
catalyses the process of PARylation by attaching the 
polymers of ADP-ribose on target protein motif via 
ester linkage and trigger the array of vital cellular 
functions viz. chromatin structure, DNA repair, trans-
criptional regulation, apoptosis, necrosis, cell separation 
and differentiation. In human, 17 members of PARP-1 
are expressed, of which NAD+ ADP-ribosyltransferase 
is associated with the nucleus and regulates at least 85% 
of the cellular NAD+ ADP-ribosyltransferase activities 
(Hur et al., 2006; Smulson et al., 2000; Corcoran et al., 
2016; Hassa et al., 2008; Phulwani et al., 2008). PARP-1 
maintains the genomic integrity by PARylation of his-
tones and other key enzymes, DNA repair mechanism, 
interprotein interactions and gene expression to ensure 
optimum cellular homeostasis (Ba et al., 2011; Chiarugi 
et al., 2003; Yu et al., 2002). However, throughout the 
cellular and/or genotoxic stress, prolong activation of 

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The objective of this study was to evaluate the role of poly (ADP-ribose) 
polymerase-1 (PARP-1) and NF-kappaB as potential risk factor in the 
development of non-small cell lung cancer in Chinese individuals (n=110) of 
either gender aged <65 years. Healthy Chinese individuals (n=110) were 
included as control. Polymerase chain reaction was used to determine the role 
of PART-1 and NF-kappaB using plasma sample. The individuals with the 
GA genotype and carrier of G allele were at high-risk of non-small cell lung 
cancer. There was involvement of dL/is of the NF-kappaB in non-small cell 
lung cancer. Our study result suggests that the PARP-1 and NF-kappaB play 
an important role in development of non-small cell lung cancer in China.  

$UWLFOH�,QIR 

Received:  17 September 2017 
Accepted:  22 September 2017 
Available Online:   30 January 2017  

 

DOI: 10.3329/bjp.v13i1.34031 
 

Cite this article: 
Chen A, Yu Z, Zhang R, Zhao Q, Ge J. 
Role of poly (ADP-ribose) polymerase
-1 and NF-kappaB as potential risk 
factor in the development of non-
small cell lung cancer in Chinese indi-
viduals. Bangladesh J Pharmacol. 
2018; 13: 30-34. 

Role of poly (ADP-ribose) polymerase-1 and NF-kappaB as 
potential risk factor in the development of non-small cell lung 
cancer in Chinese individuals 
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This work is licensed under a Creative Commons Attribution 4.0 License. You are free to copy, distribute and perform the work. You must attribute 
the work in the manner specified by the author or licensor.  



 

PARP-1 lead to decrease in β-nicotinamide adenine 
dinucleotide (NAD+) and adenosine triphosphate 
(ATP), and causes release of mitochondrial proapop-
totic protein, apoptosis-inducing factor (AIF). Such 
stress-induced dysregulation in cellular homeostasis 
mediated by PARP-1, triggers the downstream necrosis 
cascade of cell death (Ba et al., 2011; Chiarugi et al., 
2003).  

The functional role of PARP-1 and NF-kappaB in the 
non-small cell lung cancer was not evaluated among 
Chinese individuals. The objective of this study was to 
ascertain whether PARP-1 and NF-kappaB are involved 
in the development of non-small cell lung cancer 
among Chinese individuals.     

 

Materials and Methods 

Subjects 

Patients of both genders aged <65 years with a confirm-
ed diagnosis of non-small cell lung cancer (n=110) and 
healthy individuals (n=110) were recruited. The study 
protocol was approved by the Ethics Committee of the 
Anhui Provincial Hospital of the Anhui Medical Uni-
versity. The written informed consent was taken from 
each study subject. All subjects were educated about 
the study protocol and the likely benefits to the society. 
All participants undergone laboratory tests or required 
investigation to confirm their eligibility for this study.  

Assessments 

Plasma samples were collected from the study popula-
tion and the DNA from leukocytes was extracted by the 
high salt DNA extraction method. All isolated DNA 
samples were kept at -80°C for further assessment. 
Using efficient molecular biology techniques, polymer-
phisms of the PARP-1 and NFkB genes were studied in 
DNA samples by polymerase chain reaction-restriction 
fragment length polymorphism (PCR-RFLP) analysis. 
PCR-RFLP was performed using an optimized sequen-
ce of cycles: A denaturation phase at 95°C (2 min) 
followed by 30 cycles at 95°C (30 sec), 60°C (30 sec), 72°
C (1 min), and a final incubation phase at 72°C (5 min). 
The PCR products were treated with 5 U of HpyF3I 
(DdeI) (fermentas) and Bsh1236I (fermentas) at 37°C 
overnight for digestion. The resulting fragments were 
run on ethidium bromide-stained 3% agarose gel for 45 
min at 90V and quantified through direct detection 
under ultraviolet light. Likewise, the NFkB genes were 
amplified by 285 and 424 base-pair PCR fragments 
using the same reagents. The PCR running sequence 
included a denaturing step at 95°C (1 min) followed by 
35 cycles at 95°C (30 s), 61°C (30 s), 72°C (1 min) and a 
final incubation at 72°C (5 min). The PCR products 
were treated with 5 U PflMI (Van91I) and HaeIII 
(BsuRI) (Fermentas) at 37°C overnight for digestion and 

the digested products were run on ethidium bromide-
stained 3% agarose gel for 45 min at 90V and quantified 
directly under UV light.  

Statistical analysis 

Normality of data was checked using KM or SW test, 
and interpretation is solely depends on the results of p 
value of SW/KM test which confirm the normality of 
the data. Data which follow bell shaped  or Gaussian 
distribution were test using parametric test such as 
unpaired ‘t’ test for between group comparison, and if 
data not following the bell shaped or Gaussian distribu-
tion, then data were analyzed using Mann-Whitney U 
test or non-parametric test based on the number of 
treatment groups. Numerical variable was presented as 
mean (standard deviation). Qualitative outcome were 
presented as absolute number (percentage) of indivi-
duals in each category. Data from each patient was 
coded and analyzed using GraphPad Prism statistical 
analysis software (version 6.0). Quantitative variable 
was presented as mean (standard deviation), and 
analyzed by parametric/non-parametric statistical test 
based number of comparison group and distribution of 
data, using 2-sided statistical tests. Categorical variables 
were presented as absolute number and/or percentage 
of subjects in each category, and analyzed by Chi-
square or fisher exact test based on size of data, using 2-
sided statistical tests. Odd ratio was calculated using 
Chi-square or fisher exact statistical test.  

 

Results 
Demography and clinical characteristics 

The mean (SD) age of the patients of non-small cell lung 
cancer was 32.8 (2.3), whereas it was 36.5 (2.4) year in 
healthy subjects (Table I).  

Primary and secondary endpoints results 

The individuals with GA gene were at susceptible of 

 Bangladesh J Pharmacol 2018; 13: 30-34                                                                 31 

Table I 

Demographic characteristics 

Variables Patients 
(n=110) 

Healthy subjects 
(n=110) 

p 
value 

Age 32.8 
(2.3) 

36.5 
(2.4) 

>0.005 

Gender     

Men 85 80 >0.005 

Women 25 30 >0.005 

BMI 24.16 
(2.5) 

22.53 
(3.2) 

>0.005 

Data are mean (± SD) 



 

developing non-small cell lung cancer. The risk of non-
small cell lung cancer was higher in patients who had 
GA gene compared to subjects who had no GA gene 
(p<0.001). The individuals with G allele were also 

susceptible for higher risk of non-small cell lung cancer; 
risk of non-small cell lung cancer was higher in patients 
who had G allele compared to subjects who had no G 
allele (p<0.001). The individuals with C allele were 
more susceptible of non-small cell lung cancer (Table 
II). There was positive relationship of PARP-1 with non-
small cell lung cancer. Among patients of non-small cell 
lung cancer, del/ins of nuclear factor-κB1 gene was 
found associated. This suggest role of del/ins of nuclear 
factor-κB1 in progression of non-small cell lung cancer. 
Association of del/ins of NF-kB1 was observed with 
non-small cell lung cancer. However, there was no 
statistical significant association was observed among 
other genotype of nuclear factor-κB1 gene with non-
small cell lung cancer. Our finding suggested that there 
was significant association of polymorphism of PARP-1  
and nuclear factor-κB1 in patients of non-small cell lung 
cancer as compared to healthy subjects (Table II).  
                  

Discussion 
To the best of knowledge, this is the first study to 
determine involvement of PARP-1 and NF-kappaB in 
individuals with non-small cell lung cancer. In this 
study, the individuals with GA gene were at higher risk 
of non-small cell lung cancer as compared to subjects 
who had no GG gene. Also it was observed that the 
individuals with C and G allele are also susceptible of 
non-small cell lung cancer. This indicates the positive 
relationship of PARP-1 polymorphisms. In addition, 
among patients of non-small cell lung cancer, 
polymorphism of del/ins of nuclear factor-κB1 gene 
was found associated. This indicates the involvement of 
del/ins of nuclear factor-κB1 in development of non-
small cell lung cancer. The results of this study 
suggested that there was significant association of 
polymorphism of PARP-1 and nuclear factor-κB1 in 
patients of non-small cell lung cancer as compared to 
healthy subjects. Zhong et al. have demonstrated a 
novel, significant correlation between the hOGG1 
Ser326Cys polymorphism and increased lung cancer 
susceptibility in Caucasians. Authors of this study have 
indicated a need for larger-scale studies to verify the 
association of this SNP with lung cancer risk in 
Caucasians (Zhong D, 2012). Another study showed 
lymorphic variations in TP53 represent attractive 
candidate susceptibility alleles for lung cancer. In this 
study, authors have showed the accosiation of MspI 
RFLP and 16 bp duplication polymorphism in increa-
sing the risk of lung cancer (Matakidou, 2003).  

In this study, individuals with a GG genotype and 
carriers of a G allele were more susceptible to and at 
very high risk of developing non-small cell lung cancer. 
This indicates that a GA genotype might have a 
defensive influence on the disease. The possible reason 
for involvement of the GG genotype as a risk factor for 

Table II 

Genotypes involved in patients of non-small cell 
lung cancer 

Sequence Patients 
(n=110) 

Healthy 
subjects 
(n=110) 

Odd ratio 
(95% CI) 

p value 

G1672A genotype    

GG 46 43 1.5 
(0.7–3.3) 

0.32 

GA 84 46 2.4 
(1.2–3.1) 

0.004 

AA 43 42 1.5 
(0.7–2.4) 

0.63 

G1672A genotype    

G Allele 213 121 2.8 
(1.3–3.1) 

<0.001  

A Allele 27 32 

C410T genotype    

CC 43 42 1.1 
(0.3 –2.3) 

0.08 

CT 67 71 1.1 
(0.1–1.2) 

0.068 

TT 98 89 1.4 
(0.2–1.4) 

0.079 

  

C Allele 76 38 2.3 
(1.3–2.7)  

0.002  

T Allele 34 39 

NF-κB1 genotype    

Ins/ins 21 28 3.1   

del/ins 11 26 2.8 
(1.3–3.2) 

0.23 

del/del 23 21 2.2 
(2.2–2.9) 

0.19 

Allele- NF-κB1    

Ins 54 65 3.2 
(2.4–3.3)  

0.073  

Del 43 47 

NF-κBIA - genotype    

AA 56 57 1.2   

AG 89 79 3.5 
(2.4–3.6) 

0.05 

GG 76 67 2.4 
(2.4–2.2) 

0.068 

Allele involved in NF-κBIA    

AA 231 239 2.2 
(3.3–3.4)  

0.22  

AG 39 42 

Allele - C410T  

32 Bangladesh J Pharmacol 2018; 13: 30-34                                                                



 

non-small cell lung cancer could be explained by 
molecular heterosis, which is observed in approx. 50% 
of cases of gene associations (Comings, 2000). Also it 
was observed the involvement of a NF-κB1 polymer-
phism in the increased risk of non-small cell lung 
cancer. This study results showed involvement of PARP
-1 and NF-κB in non-small cell lung cancer indicates 
PARP-1 and NF-κB as a promising therapeutic target in 
clinical treatment of non-small cell lung cancer. In 
addition, previous study demonstrated that PARP-1 
inhibitors inhibit apoptotic cell death in neuronal tissue 
by preventing the activation of the apoptosis pathway 
(Yu, 2005). In addition, reports indicate that PARP-1 
inhibitors have strong anti-inflammatory properties 
resulting in reduced 1) polymorphonuclear leukocyte 
infiltration, 2) activation of NF-κB, and 3) degree of 
tissue injury (Comings, 2000; Yu, 2005; Chiarugi A 2002; 
Scott GS, 1999). Differential expression of the desired 
gene product in the target tissue is central to the con-
cept of gene therapy (Trofimova, 2002). The develop-
ment of potent PARP-1 inhibitors has become of 
increasing clinical interest in various disease conditions. 
This study results suggest that the PARP inhibitor-
based gene therapy as a new treatment paradigm for 
non-small cell lung cancer. This study results provide 
proof-of-principle for a novel therapeutic strategy for 
the treatment of non-small cell lung cancer. 

This was the first study to suggest the association of 2 
single nucleotide polymorphisms (SNPs) of PARP-1 
(C410T and G1672A) with non-small cell lung cancer. 
The involvement of these SNPs in non-small cell lung 
cancer has not yet been established. Since PARP-1  
catalyzes poly (ADP-ribosyl)ation to various proteins 
involved in many cellular processes, including DNA 
damage detection and repair, and cell proliferation and 
death (Yu, 2002). Several reports showed the involve-
ment of polymorphisms of PARP-1 in susceptibility of 
developing various diseases such as cancer, cataract 
(Cui, 2017)  and Graves’ disease (Niyazoglu, 2014). 
Thus, finding of this study encourages the researchers 
to focus on two SNPs of PARP-1 (C410T and G1672A) 
in other disease conditions. Since the study was 
designed as pilot study and conducted at single study 
center in China (limitation of study). Therefore, the 
present findings cannot be generalized to the overall 
Chinese population. Based on the study results, study 
results encourage for conducting large multi-centric 
randomized clinical study in future to generalize the 
findings of this study. The hypothesis of this study for 
relationship between PARP-1 and NFkB with non-small 
cell lung cancer was met. Overall, results showed the 
polymorphism of PARP-1 and nuclear factor-κB1 is 
involved in development of non-small cell lung cancer 
in Chinese individuals   

Conclusion 

The study result showed the involvement of PARP-1  
and NFkB in development of non-small cell lung cancer 
among Chinese individuals.   

 

Acknowledgement 
All authors would like to thank the subjects for their 
participation in this study.  

 

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