Journal of Carcinogenesis

ORIGINAL ARTICLE
Year
: 2020  |  Volume : 19  |  Issue : 1  |  Page : 11-

Identification of suitable reference genes in blood samples of carcinoma lung patients using quantitative real-time polymerase chain reaction


Shashi Ranjan Mani Yadav1, Bela Goyal1, Raman Kumar1, Sweety Gupta2, Amit Gupta3, Anissa Atif Mirza1, Gaurav Sharma2, Shalinee Rao4, Rajesh Pasricha2, Manoj Gupta2 
1 Department of Biochemistry, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
2 Department of Radiation Oncology, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
3 Department of Surgery, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India
4 Department of Pathology and Laboratory Medicine, All India Institute of Medical Sciences, Rishikesh, Uttarakhand, India

Correspondence Address:
Sweety Gupta
Department of Radiation Oncology, All India Institute of Medical Sciences, Rishikesh - 249 203, Uttarakhand
India

INTRODUCTION: Lung cancer (LC), among all other cancers, is the leading cause of death worldwide, while the third most common cancer-causing mortality in India. Several techniques of the assay for early detection of cancer that improve survival rates have been employed in tissues and cell lines. Reverse transcriptase quantitative polymerase chain reaction (RTqPCR) is one of the most common techniques employed for gene expression studies for the normalization of a target gene using a reference gene (RG). The present study used the three most common RGs: glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-Actin, and 18s ribosomal ribonucleic acid (18s rRNA), which were assessed by qPCR to validate, as of which is a more effective RG in blood samples of LC patients. MATERIALS AND METHODS: A total of thirty participants with LC of non-small cell and small cell type were included along with twenty healthy controls. Ribonucleic acid (RNA) isolated from peripheral blood mononuclear cells was quantified, prepared for complementary deoxyribose nucleic acid synthesis, and analyzed for expression of three RG on RTqPCR. RESULTS: Expression levels as Ct values of studied RG were reported as mean ± standard deviation for GAPDH (26.97 ± 5.107), β-actin (20.5 ± 2.3), and 18s rRNA (25.10 ± 4.075). GAPDH showed the lowest expression, whereas β-actin showed the highest expression among the studied RG in subjects of LC. The expression of GAPDH and 18s rRNA were statistically significantly lower than β-actin (p < 0.0001), whereas expression levels of GAPDH and 18s rRNA were comparable. However, the expression level of only β-actin in LC patients was comparable with healthy controls with P < 0.1611 at 95% confidence interval. CONCLUSION: It is concluded that β -actin may be considered the most suitable RG isolated and studied from peripheral blood mononuclear cells using RT qPCR in LC.


How to cite this article:
Yadav SR, Goyal B, Kumar R, Gupta S, Gupta A, Mirza AA, Sharma G, Rao S, Pasricha R, Gupta M. Identification of suitable reference genes in blood samples of carcinoma lung patients using quantitative real-time polymerase chain reaction.J Carcinog 2020;19:11-11


How to cite this URL:
Yadav SR, Goyal B, Kumar R, Gupta S, Gupta A, Mirza AA, Sharma G, Rao S, Pasricha R, Gupta M. Identification of suitable reference genes in blood samples of carcinoma lung patients using quantitative real-time polymerase chain reaction. J Carcinog [serial online] 2020 [cited 2020 Nov 28 ];19:11-11
Available from: http://www.carcinogenesis.com/article.asp?issn=1477-3163;year=2020;volume=19;issue=1;spage=11;epage=11;aulast=Yadav;type=0