Amygdalin inhibits the growth of renal cell carcinoma cells in vitro

Juengel,Eva; Thomas,Anita; Rutz,Jochen; Makarevic,Jasmina; Tsaur,Igor; Nelson,Karen; Haferkamp,Axel; Blaheta,Roman  A.

doi:10.3892/ijmm.2015.2439

Amygdalin inhibits the growth of renal cell carcinoma cells in vitro

Authors:
- Eva Juengel
- Anita Thomas
- Jochen Rutz
- Jasmina Makarevic
- Igor Tsaur
- Karen Nelson
- Axel Haferkamp
- Roman A. Blaheta
View Affiliations

Affiliations: Department of Urology, Goethe-University Hospital, D-60590 Frankfurt am Main, Germany, Department of Vascular and Endovascular Surgery, Goethe-University Hospital, D-60590 Frankfurt am Main, Germany
Published online on: December 21, 2015 https://doi.org/10.3892/ijmm.2015.2439
Pages: 526-532

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Although amygdalin is used by many cancer patients as an antitumor agent, there is a lack of information on the efficacy and toxicity of this natural compound. In the present study, the inhibitory effect of amygdalin on the growth of renal cell carcinoma (RCC) cells was examined. Amygdalin (10 mg/ml) was applied to the RCC cell lines, Caki-1, KTC-26 and A498, for 24 h or 2 weeks. Untreated cells served as controls. Tumor cell growth and proliferation were determined using MTT and BrdU tests, and cell cycle phases were evaluated. Expression of the cell cycle activating proteins cdk1, cdk2, cdk4, cyclin A, cyclin B, cyclin D1 and D3 as well as of the cell cycle inhibiting proteins p19 and p27 was examined by western blot analysis. Surface expression of the differentiation markers E- and N-cadherin was also investigated. Functional blockade by siRNA was used to determine the impact of several proteins on tumor cell growth. Amygdalin treatment caused a significant reduction in RCC cell growth and proliferation. This effect was correlated with a reduced percentage of G2/M-phase RCC cells and an increased percentage of cells in the G0/1-phase (Caki-1 and A498) or cell cycle arrest in the S-phase (KTC-26). Furthermore, amygdalin induced a marked decrease in cell cycle activating proteins, in particular cdk1 and cyclin B. Functional blocking of cdk1 and cyclin B resulted in significantly diminished tumor cell growth in all three RCC cell lines. Aside from its inhibitory effects on growth, amygdalin also modulated the differentiation markers, E- and N-cadherin. Hence, exposing RCC cells to amygdalin inhibited cell cycle progression and tumor cell growth by impairing cdk1 and cyclin B expression. Moreover, we noted that amygdalin affected differentiation markers. Thus, we suggest that amygdalin exerted RCC antitumor effects in vitro.

View Figures

View References

1	Maute L, Grünwald V, Weikert S, Kube U, Gauler T, Kahl C, Burkholder I and Bergmann L: Therapy of mRCC beyond mTOR-inhibition in clinical practice: results of a retrospective analysis. J Cancer Res Clin Oncol. 140:823–827. 2014. View Article : Google Scholar : PubMed/NCBI
2	Najjar YG and Rini BI: Novel agents in renal carcinoma: a reality check. Ther Adv Med Oncol. 4:183–194. 2012. View Article : Google Scholar : PubMed/NCBI
3	Sun M, Thuret R, Abdollah F, Lughezzani G, Schmitges J, Tian Z, Shariat SF, Montorsi F, Patard JJ, Perrotte P and Karakiewicz PI: Age-adjusted incidence, mortality, and survival rates of stage-specific renal cell carcinoma in North America: a trend analysis. Eur Urol. 59:135–141. 2011. View Article : Google Scholar
4	Huebner J, Micke O, Muecke R, Buentzel J, Prott FJ, Kleeberg U and Senf B: User rate of complementary and alternative medicine (CAM) of patients visiting a counseling facility for CAM of a German comprehensive cancer center. Anticancer Res. 34:943–948. 2014.PubMed/NCBI
5	Saghatchian M, Bihan C, Chenailler C, Mazouni C, Dauchy S and Delaloge S: Exploring frontiers: use of complementary and alternative medicine among patients with early-stage breast cancer. Breast. 23:279–285. 2014. View Article : Google Scholar : PubMed/NCBI
6	Bolarinwa IF, Orfila C and Morgan MR: Determination of amygdalin in apple seeds, fresh apples and processed apple juices. Food Chem. 170:437–442. 2015. View Article : Google Scholar
7	Tanaka R, Nitta A and Nagatsu A: Application of a quantitative 1H-NMR method for the determination of amygdalin in Persicae semen, Armeniacae semen, and Mume fructus. J Nat Med. 68:225–230. 2014. View Article : Google Scholar
8	Lee J, Zhang G, Wood E, Rogel Castillo C and Mitchell AE: Quantification of amygdalin in nonbitter, semibitter, and bitter almonds (Prunus dulcis) by UHPLC-(ESI)QqQ MS/MS. J Agric Food Chem. 61:7754–7759. 2013. View Article : Google Scholar : PubMed/NCBI
9	Moss RW: The laetrile controversy. The Cancer Industry. The Classic Expose on the Cancer Establishment. First Equinox Press; Brooklyn, NY: pp. 131–152. 1996
10	Curt GA: Unsound methods of cancer treatment. Princ Pract Oncol Updates. 4:1–10. 1990.
11	PDQ Cancer Complementary and Alternative Medicine Editorial Board Laetrile/Amygdalin (PDQ®): Health Professional Version PDQ Cancer Information Summaries [Internet]. Bethesda (MD): National Cancer Institute (US); 2002–2015
12	Moss RW: Patient perspectives: Tijuana cancer clinics in the post-NAFTA era. Integr Cancer Ther. 4:65–86. 2005. View Article : Google Scholar : PubMed/NCBI
13	Moertel CG, Fleming TR, Rubin J, Kvols LK, Sarna G, Koch R, Currie VE, Young CW, Jones SE and Davignon JP: A clinical trial of amygdalin (Laetrile) in the treatment of human cancer. N Engl J Med. 306:201–206. 1982. View Article : Google Scholar : PubMed/NCBI
14	Moertel CG, Ames MM, Kovach JS, Moyer TP, Rubin JR and Tinker JH: A pharmacologic and toxicological study of amygdalin. JAMA. 245:591–594. 1981. View Article : Google Scholar : PubMed/NCBI
15	Ames MM, Moyer TP, Kovach JS, Moertel CG and Rubin J: Pharmacology of amygdalin (laetrile) in cancer patients. Cancer Chemother Pharmacol. 6:51–57. 1981.PubMed/NCBI
16	Newell GR and Ellison NM: Ethics and designs: laetrile trials as an example. Cancer Treat Rep. 64:363–365. 1980.PubMed/NCBI
17	Wahab MF, Breitbach ZS, Armstrong DW, Strattan R and Berthod A: Problems and pitfalls in the analysis of amygdalin and its epimer. J Agric Food Chem. 63:8966–8973. 2015. View Article : Google Scholar : PubMed/NCBI
18	Makarević J, Rutz J, Juengel E, Kaulfuss S, Reiter M, Tsaur I, Bartsch G, Haferkamp A and Blaheta RA: Amygdalin blocks bladder cancer cell growth in vitro by diminishing cyclin A and cdk2. PLoS One. 9:e1055902014. View Article : Google Scholar
19	Qian L, Xie B, Wang Y and Qian J: Amygdalin-mediated inhibition of non-small cell lung cancer cell invasion in vitro. Int J Clin Exp Pathol. 8:5363–5370. 2015.PubMed/NCBI
20	Chen Y, Ma J, Wang F, Hu J, Cui A, Wei C, Yang Q and Li F: Amygdalin induces apoptosis in human cervical cancer cell line HeLa cells. Immunopharmacol Immunotoxicol. 35:43–51. 2013. View Article : Google Scholar
21	Chang HK, Shin MS, Yang HY, Lee JW, Kim YS, Lee MH, Kim J, Kim KH and Kim CJ: Amygdalin induces apoptosis through regulation of Bax and Bcl-2 expressions in human DU145 and LNCaP prostate cancer cells. Biol Pharm Bull. 29:1597–1602. 2006. View Article : Google Scholar : PubMed/NCBI
22	Chang WL, Yu CC, Chen CS and Guh JH: Tubulin-binding agents down-regulate matrix metalloproteinase-2 and -9 in human hormone-refractory prostate cancer cells - a critical role of Cdk1 in mitotic entry. Biochem Pharmacol. 94:12–21. 2015. View Article : Google Scholar : PubMed/NCBI
23	Visconti R, Della Monica R, Palazzo L, D'Alessio F, Raia M, Improta S, Villa MR, Del Vecchio L and Grieco D: The Fcp1-Wee1-Cdk1 axis affects spindle assembly checkpoint robustness and sensitivity to antimicrotubule cancer drugs. Cell Death Differ. 22:1551–1560. 2015. View Article : Google Scholar : PubMed/NCBI
24	Wang WT, Catto JW and Meuth M: Differential response of normal and malignant urothelial cells to CHK1 and ATM inhibitors. Oncogene. 34:2887–2896. 2015. View Article : Google Scholar
25	Carcagno AL, Marazita MC, Ogara MF, Ceruti JM, Sonzogni SV, Scassa ME, Giono LE and Cánepa ET: E2F1-mediated upregulation of p19INK4d determines its periodic expression during cell cycle and regulates cellular proliferation. PLoS One. 6:e219382011. View Article : Google Scholar : PubMed/NCBI
26	Yuan H, Meng X, Guo W, Cai P, Li W, Li Q, Wang W, Sun Y, Xu Q and Gu Y: Transmembrane-bound IL-15-promoted epithelial-mesenchymal transition in renal cancer cells requires the Src-dependent Akt/GSK-3β/β-catenin pathway. Neoplasia. 17:410–420. 2015. View Article : Google Scholar : PubMed/NCBI
27	He H and Magi-Galluzzi C: Epithelial-to-mesenchymal transition in renal neoplasms. Adv Anat Pathol. 21:174–180. 2014. View Article : Google Scholar : PubMed/NCBI
28	Zhang X, Ren J, Yan L, Tang Y, Zhang W, Li D, Zang Y, Kong F and Xu Z: Cytoplasmic expression of pontin in renal cell carcinoma correlates with tumor invasion, metastasis and patients' survival. PLoS One. 10:e01186592015. View Article : Google Scholar : PubMed/NCBI
29	Shimazui T, Kojima T, Onozawa M, Suzuki M, Asano T and Akaza H: Expression profile of N-cadherin differs from other classical cadherins as a prognostic marker in renal cell carcinoma. Oncol Rep. 15:1181–1184. 2006.PubMed/NCBI
30	Cheng C, Wan F, Liu L, Zeng F, Xing S, Wu X, Chen X and Zhu Z: Overexpression of SATB1 is associated with biologic behavior in human renal cell carcinoma. PLoS One. 9:e974062014. View Article : Google Scholar : PubMed/NCBI
31	Huang J, Yao X, Zhang J, Dong B, Chen Q, Xue W, Liu D and Huang Y: Hypoxia-induced downregulation of miR-30c promotes epithelial-mesenchymal transition in human renal cell carcinoma. Cancer Sci. 104:1609–1617. 2013. View Article : Google Scholar : PubMed/NCBI
32	Gervais ML, Henry PC, Saravanan A, Burry TN, Gallie BL, Jewett MA, Hill RP, Evans AJ and Ohh M: Nuclear E-cadherin and VHL immunoreactivity are prognostic indicators of clear-cell renal cell carcinoma. Lab Invest. 87:1252–1264. 2007. View Article : Google Scholar : PubMed/NCBI
33	Ho MY, Tang SJ, Chuang MJ, Cha TL, Li JY, Sun GH and Sun KH: TNF-α induces epithelial-mesenchymal transition of renal cell carcinoma cells via a GSK3β-dependent mechanism. Mol Cancer Res. 10:1109–1119. 2012. View Article : Google Scholar : PubMed/NCBI
34	Harada K, Miyake H, Kusuda Y and Fujisawa M: Expression of epithelial-mesenchymal transition markers in renal cell carcinoma: impact on prognostic outcomes in patients undergoing radical nephrectomy. BJU Int. 110:E1131–E1137. 2012. View Article : Google Scholar : PubMed/NCBI