In vitro antitumor efficacy of Kochia indica extract on human hepatocellular carcinoma cell line with or without 5-fluorouracil

Abstract

Aim: Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and the sixth most common cancer worldwide. The resistance to chemotherapy is a major obstacle in the treatment of HCC, necessitating the discovery of additional agents. There is a growing use of anticancer complementary and alternative medicine worldwide. Therefore, the aim of present study was focused on the confirmation of the suitability and validity of the new markers which would be achieved by demonstrating their significant change and reproducible expression during disease and disease management.

Methods: HepG₂ cell line was used to provide a source for HCC cells. The cell cultures were divided into 4 groups: control untreated group, 5-fluorouracil (5-FU) treated group as a standard chemotherapy for HCC (positive control) with the following doses (15.625, 31.2, 62.5, 125, 250 µg/mL), Kochia indica extract treated group with the following concentration (12.5, 25, 50, 100, 200 µg/mL) and the group treated with a combination of 5-FU and Kochia indica in different ratios.

Results: Treatment with Kochia indica extract, 5-FU and the combined treatment showed a significant cytotoxicity to HepG₂ cells, with different IC₅₀ values, when compared to the control. Regarding toxic effect, 5-FU showed IC₅₀ = 237.56 µg/mL which is lower cytotoxic in compared to Kochia indica with IC₅₀ =120.5 µg/mL. The results also revealed that tumor cells were more resistant to 5-FU. Alternatively, the co-treatment with Kochia indica extract ameliorated the toxicity induced by 5-FU and enhanced its therapeutic potency, either by synergistic effect of both agents and/or due to its flavonoid components that may enhance the physiological properties of the cell membranes, facilitating 5-FU entrance into tumor cells. This decreased its therapeutic dose to less than 250 µg/mL by combination therapy.

Conclusion: Present findings assume that Kochia indica extract co-therapy can ameliorate the side effects of 5-FU on HepG₂ by enhancing its cellular uptake.

Keywords

Hepatocellular carcinoma / Kochia indica / 5-fluorouracil / cells of hepatoma cell line / chemosensitization

References

[1] Ferlay J,Dikshit R,Mathers C,Parkin DM,Bray F.Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012..Int J Cancer2015;136:E359-86

[2] Dhanasekaran R,Torbenson MS.Clinical implications of basic research in hepatocellular carcinoma..J Hepatol2016;64:736-45 PMCID:PMC5039166

[3] Colquhoun SD.Hepatocellular carcinoma: the current role of surgical intervention..Crit Rev Oncog2016;21:93-103

[4] Shaker MK,Khalifa MO.Epidemiological characteristics of hepatocellular carcinoma in Egypt: a retrospective analysis of 1313 cases..Liver Int2013;33:1601-6

[5] Mazzio EA.In vitro screening for the tumoricidal properties of international medicinal herbs..Phytother Res2009;23:385-98 PMCID:PMC2650746

[6] Abdel-Hamid NM,Abdel-Bakey AI.Polyol profile as an early diagnostic and prognostic marker in natural product chemoprevention of hepatocellular carcinoma in diabetic rats..Diabetes Res Clin Pract2011;92:228-37

[7] El Shereef AA.Kochiaplant as potential forage for ruminants under desert conditions..Annu Res Rev Bio2016;10:1-6

[8] Haroun SA.Mutageneic effects of Kochiaindica extract on Viciafaba L..J Am Sci2010;6:292-7

[9] Youssef RSA.Medicinal and non-medicinal uses of some plants found in the middle region of Saudi Arabia..J Med Plant Res2013;7:2501-17

[10] Khare CP.Indian Medicinal Plants: An Illustrated Dictionary..2007;New YorkSpringer812

[11] Nasr M,Abdelazem A.In vitro and in vivo evaluation of cubosomes containing 5-fluorouracil for liver targeting..Acta Pharm Sin B2015;5:79-88 PMCID:PMC4629209

[12] Prayong P,Weerapreeyakul N.Cytotoxic activity screening of some indigenous Thai plants..Fitoterapia2008;79:598-601

[13] Van der BliekAM AM,Ten Houte de Lange T,Van der Velde-Koerts T.The human mdr3 gene encodes a novel p-glycoprotein homologue and gives rise to alternatively spliced mRNAs in liver..EMBO J1987;6:3325-31

[14] Fotakis G.In vitro cytotoxicity assays: comparison of LDH, neutral red, MTT and protein assay in hepatoma cell lines following exposure to cadmium chloride..Toxicol Lett2006;160:171-7

[15] Machana S,Barusrux S,Sripanidkulchai B.Cytotoxic and apoptotic effects of six herbal plants against the human hepatocarcinoma (HepG2) cell line..Chin Med2011;6:39 PMCID:PMC3224580

[16] Chou TC.Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors..Adv Enzyme Regul1984;22:27-55

[17] AbouZid S.Evaluation of usefulness of wild plants growing in Beni-Sueif Region, Upper Egypt..Funct Plant Sci Biotechnol2011;5:91-3

[18] Marelli L,Triantos C,Cholongitas E,Burroughs AK.Treatment outcomes for hepatocellular carcinoma using chemoembolization in combination with other therapies..Cancer Treat Rev2006;32:594-606

[19] Abdel-Hamid NM,Mahmoud AW,Youssof M.A survey on herbal management of hepatocellular carcinoma..World J Hepatol2011;3:175-83 PMCID:PMC3158906

[20] Yoo BK,Vozhilla N,Chen D,Shah K,Hansen U,Sarkar D.Identification of genes conferring resistance to 5-fluorouracil..Proc Natl Acad Sci U S A2009;106:12938-43 PMCID:PMC2722317

[21] Aboul-Enein AM,Shalaby EA.Traditional medicinal plants research in Egypt: studies of antioxidant and anticancer activities..J Med Plant Res2012;6:689-703

[22] Holaha NS,Aiada HA.Hepatocellular carcinoma in Egypt: epidemiological and histopathological properties..Menoufia Med J2015;28:718-24