Genetic Analysis of Restriction Fragment Length Polymorphism of TLL1 Gene (rs17047200) in Patients of Hepatocellular Carcinoma

RFLP of TLL1 gene


  • Saima Sharif Department of Zoology, Lahore College for Women University, Lahore
  • Sadia Ashraf Department of Zoology, Lahore College for Women University, Lahore
  • Shagufta Naz Department of Zoology, Lahore College for Women University, Lahore
  • Sher Zaman Saf Interdisciplinary Research Centre in Biomedical Materials, COMSATS University, Lahore
  • Muhammad Saqib District Head Quarter Hospital, Nankana Sahib, Sheikhpura



Liver Cancer, Polymorphism, Hepatocellular Carcinoma, PCR, TLL1 gene


To find out the association of polymorphism of the TLL1 gene in hepatocellular carcinoma. A cross-sectional study was conducted from January 2020 to September 2020. Subjects were enrolled from Mayo Hospital, Lahore, Jinnah Hospital Lahore and the Liver Transplant Unit of Sheikh Zayed Hospital, Lahore. A total of 200 individuals were registered and segregated into the Control group (n=100) and hepatocellular carcinoma (HCC) group (n=100). DNA was extracted from obtained blood samples and Restriction Fragments Length Polymorphism (RFLP) was carried out at the laboratories of LCWU by using specific primers and restriction endonuclease enzymes. The data were analyzed statistically. The high proportion of smoking, hepatitis B, hepatitis C, cirrhosis and Body Mass Index (BMI) were established risk factors in the HCC group. Subjects with hepatocellular carcinoma had low socioeconomic status. Heterozygous bands in the HCC group were observed after RFLP. TLL1 genotype was AA (72 %) and AT/TT
(28 %). The patient’s clinical aspects were similar across TLL1 genotypes. It was concluded that RFLP on the exon region by using their specific enzymes HpyCH4III showed heterozygous bands in the HCC group that indicated a mutation in the TLL1 gene though this mutation does have a significant association with HCC.


F. Bray, J. Ferlay, I. Soerjomataram, R. L. Siegel, L.A. Torre, and A. Jemal. Global cancer statistics: 2018 GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. A Cancer Journal for Clinicians 68(6): 394-424 (2018).

J.L. Petrick, A.A. Florio, A. Znaor, D. Ruggieri, M. Laversanne, C.S. Alvarez, J. Ferlay, P.C. Valery, F. Bray, and K.A. McGlynn. International trends in hepatocellular carcinoma incidence, 1978–2012. International Journal of Cancer 147(2): 317-30 (2020).

H.B. El-Serag. Epidemiology of viral hepatitis and hepatocellular carcinoma. Gastroenterology 142(6): 1264-73 (2012).

A. Tang, O. Hallouch, V. Chernyak, A. Kamaya, and C.B. Sirlin. Epidemiology of hepatocellular carcinoma: target population for surveillance and diagnosis. Abdominal Radiology 43(1): 13- 25(2018).

H.K. Lyerly, M.R. Fawzy, Z. Aziz, R. Nair, C.S. Pramesh, V. Parmer, P.M. Parikh, R. Jamal, A. Irumnaz, J. Ren, and M.R. Stockler. Regional variation in identified cancer care needs of early career oncologists in China, India, and Pakistan. The Oncologist 20(5): 532 (2015).

J. Gao, L. Xie, W.S. Yang, W. Zhang, S. Gao, J. Wang, and Y.B. Xiang. Risk factors of hepatocellular carcinoma-current status and perspectives. Asian Pacific Journal of Cancer Prevention, 13(3): 743- 52 (2012).

A.B. Hafeez Bhatti, F.S. Dar, A. Waheed, K. Shafique, F. Sultan, and N.H. Shah. Hepatocellular carcinoma in Pakistan: national trends and global perspective. Gastroenterology Research and Practice 3(1) (2016).

L. Sieron, M. Lesiak, I. Schisler, Z. Drzazga, A. Fertala and A.L. Sieron. Functional and structural studies of tolloid-like 1 mutants associated with atrial-septal defect. Bioscience Reports 39(1) (2019).

A. Kiso, Y. Toba, S. Tsutsumi, S. Deguchi, K. Igai, S. Koshino, Y. Tanaka, K. Takayama, and H. Mizuguchi. Tolloid‐Like 1 Negatively Regulates Hepatic Differentiation of Human Induced Pluripotent Stem Cells Through Transforming Growth Factor β Signaling. Hepatology Communications 4(2): 255-67 (2020).

J. Grimberg, J. Nawoschik, L. Belluscio, R. McKee, A. Turck, and A. Eisenberg. A simple and efficient non-organic procedure for the isolation of genomic DNA from blood. Nucleic Acids Research 17 (20): 8390 (1989).

R. Siegel, C. DeSantis, K. Virgo, K. Stein, A. Mariotto, T. Smith, D. Cooper, T. Gansler, C. Lerro, S. Fedewa, and C. Lin. Cancer treatment and survivorship statistics, 2012. CA: A Cancer Journal for Clinicians 62(4): 220-241 (2012).

V.V. Padma. An overview of targeted cancer therapy. BioMedicine 5(4): 1-6 (2015).

J. Bruix, W.Y. Tak, A. Gasbarrini, A. Santoro, M. Colombo, H.Y. Lim, V. Mazzaferro, R. Wiest, M. Reig, A. Wagner, and L. Bolondi. Regorafenib as second-line therapy for intermediate or advanced hepatocellular carcinoma: multicentre, open-label, phase II safety study. European Journal of Cancer 49(16): 3412-9 (2013).

K.P. Weber, S.T. Aw, M.J. Todd, L.A. McGarvie, S. Pratap, I.S. Curthoys, and G. M. Halmagyi. Inter RFLP of TLL1 gene 69 ocular differences of the horizontal vestibulo-ocular reflex during impulsive testing. Progress in Brain Research, 1(171): 195-8 (2008).

D. Liu, A. Ahmet, L. Ward, P. Krishnamoorthy, E.D. Mandelcorn, R. Leigh, J.P. Brown, A. Cohen, and H. Kim. A practical guide to the monitoring and management of the complications of systemic corticosteroid therapy. Allergy, Asthma and Clinical Immunology 9(1): 1-25 (2013).

H.G. Li, F.F. Liu, H. Q. Zhu, X. Zhou, J. Lu, H. Chang, and J.H. Hu. Association of PTEN gene polymorphisms with liver cancer risk. International Journal of Clinical and Experimental Pathology 8(11): 15198 (2015).

K. Matsuura, H. Sawai, K. Ikeo, S. Ogawa, E. Iio, M. Isogawa, N. Shimada, A. Komori, H. Toyoda, T. Kumada, and T. Namisaki. Genome-wide association study identifies TLL1 variant associated with development of hepatocellular carcinoma after eradication of hepatitis C virus infection. Gastroenterology 152(6): 1383-94 (2017).




How to Cite

Saima Sharif, Sadia Ashraf, Shagufta Naz, Sher Zaman Saf, & Muhammad Saqib. (2022). Genetic Analysis of Restriction Fragment Length Polymorphism of TLL1 Gene (rs17047200) in Patients of Hepatocellular Carcinoma: RFLP of TLL1 gene. Proceedings of the Pakistan Academy of Sciences: Part B (Life and Environmental Sciences), 59(2), 65–70.