Fig. 2.
Antiproliferative effects of HCCMF in vivo. a, Mice were exposed to either HCCMF exposure with a xenograft-specific SAR of 67 mW/kg or are assigned to the control treatment group 3 h per day. Control group is comprised of mice receiving either randomly chosen frequencies (RCF) with a xenograft-specific SAR of 67 mW/kg and those receiving no exposure (SHAM). Tumour volume is measured three times per week and volume is calculated as (Length × (Width)2) / 2. After six weeks of exposure, statistical significance between treated and control groups has been achieved; [Week five (p = 0·045), Week six (p = 0·019) Student's two-tailed t-test] and [Test for treatment by time interaction (p < 0·001)]. b, Sequential exposure to randomly chosen and HCCMF. Five mice carrying Huh7 xenografts were exposed to RCF (grey line) 3 h per day for ten weeks during which the average tumour volume increased by 48%. Exposure was switched to HCCMF (blue line) after ten weeks. Exposure to EMF was discontinued for one week after four weeks exposure, then resumed. Mice were sacrificed at the end of week 16. c, Patient-derived HCC xenografts (PDX) exposed to HCCMF or not exposed to EMF. Patient derived xenografts from a 63-year-old male with hepatocellular carcinoma. Mice received either HCCMF exposure (HCCMF; N = 6) or received no treatment (SHAM; N = 4). Tumour volume measured three times per week and volume is calculated as (Length x Width2) / 2. After eight weeks of exposure, statistical significance had been achieved; [Week four (p = 0·0176), Week five (p = 0·0211) Student's two-tailed t-test]. At week six, all mice in the Sham group expired and tumour volume was imputed, Week six (p = 0·0553) [student's t-test]. [Test for treatment by time interaction (p = 0·0006)]. d, Ki-67 staining of SHAM, RCF and HCCMF treated tumours. [Anova: F = (2, 33) 67.55, p ≤.0001]. [Post-Hoc Tukey Test: Sham vs RCF p = 0·2067, SHAM vs HCCMF p < 0·0001, RCF vs HCCMF p < 0·0001.] e, Cyclin D1 staining of SHAM, RCF and HCCMF treated tumours. [Anova: F = (2, 33) 23·29, p < 0·0001]. [Post-Hoc Tukey test: Sham vs RCF p = 0·1379, SHAM vs HCCMF p < 0·0001, RCF vs HCCMF p = 0·0005]. f, p21 staining of SHAM, RCF and HCCMF treated tumours [Anova: F = (2, 33) 6·907, p = 0·0031]. [Post-Hoc Tukey test: Sham vs RCF p = 0·7373, Sham vs HCCMF p = 0·0411, RCF vs HCCMF p = 0·0049]. g, IHC of Huh-7 Xenograft tumours and all images at 20X (Lens; scale bar is 50 um). Tumour and intestinal crypt cell proliferation in mice exposed to HCCMF, RCF, and mice not exposed to EMF. Representative figures and graphs (d-f) represent mean +/− SEM (SHAM: N = 3; RCF: N = 3; HCCMF: N = 6) and three randomly selected fields of view per slide were used to quantify all staining. BRDU staining of SHAM, RCF and HCCMF treated tumours showed positive staining in all crypts; no statistics performed. (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)