Abstract
Cervical cancer (CC) is one of the most common cancers in the world, considered to be a direct result from infection with the Human papillomavirus (HPV). Other possible contributing factors towards CC include multiple sexual partners, having sexual activity from a very early age and immunodeficiency. In 2014, it was concluded that 19 098 women in South Africa died as a result of CC. Another report noted that CC is the most common form of cancer amongst black South African women, particularly those over the age of 35 years. Rubus idaeus (commonly referred to as red raspberry), is a plant used most commonly to treat disorders of the female genital tract. It is said to possess high antioxidant properties and has shown many cytotoxic properties in cases of cancer in recent studies. Low level laser therapy (LLLT) makes use of specific wavelengths and fluence to repair damaged tissue. Some in vivo studies done previously suggest that LLLT has the ability to induce apoptosis in cancer cells. No research demonstrating the relationship between Rubus idaeus (R. idaeus) and LLLT on CC has been conducted to date. The aim of this study was to compare the in vitro effects of R. idaeus (Homeopathic mother tincture, D6 and 30cH) and LLLT on CC cell lines. This quantitative in vitro research study took place at the Laser Research Centre of the University of Johannesburg. This study made use of HeLa cells (ATCC ccl-2) and R. idaeus (D3, D6 and 30 cH Homeopathic preparations) and LLLT at 680 nm at 5, 10 and 15 J/cm2 doses. A HeLa cell culture (ATCC CCL-2) was cultured in Dulbecco’s Modified Eagle Medium and sub-cultured from 75 cm2 flasks into 3.3 cm diameter culture plates for experimentation. There were three cell groups in this study with each experiment repeated a total of 3 times in duplicate. Each of the three experimental cell groups were divided as follows; untreated control cells (CC cells not exposed to R. idaeus or LLLT), CC cells treated with R. idaeus (D3, D6 and 30cH), CC cells treated with a combination of the optimal performing potency of R. idaeus and LLLT at 680 nm diode lasers with three fluencies of 5, 10 and 15 J/cm2. The effectiveness of these treatments were measured as follows; cell-viability was measured by using Trypan Blue assay, morphology by inverted light microscope and cytotoxicity was measured by observing the different levels of Lactate Dehydrogenase (LDH) present in the culture medium. Apoptosis of the HeLa cell culture was analyzed via Annexin V/PI and Hoechst stain and iv proliferation was observed by measuring Adenosine Triphosphate (ATP) levels in the cells. The experimental procedure was designed as a means to determine if there was a difference in the outcome of CC cell death if the cells were exposed to a single treatment with various potencies of R. idaeus or a combined treatment with the optimal potency of R. idaeus (D3) with laser 680 nm after a 24 h interval. The single and combined treatment protocol was completed a total of 6 times (n=6) for all experiments except for Annexin V/PI staining where the experiments were completed a total of 3 times (n=3). The data for both experimental procedures were analysed by using Sigma Plot version 14.0 computer software. The t-test was used in order to examine the effects of the various treatment protocols in inducing cancer cell death. In each instance, a statistical difference of p<0.05 was identified. Morphological changes indicated that the single treatment application of R. idaeus D3 had the ability to induce CC cell death in vitro however this was not the case in the combined treatment protocol of R. idaeus D3 and LLLT 680 nm. The combined treatment protocol demonstrated a photobiomodulatory effect which enhanced cancer cell growth. Based on statistical analysis, the results demonstrated that a single treatment protocol of R. idaeus D3 increased apoptosis of CC cells in vitro. However, more research needs to be conducted in order to examine the systemic effects of this remedy preparation.
M.Tech. (Homoeopathy)