Enhancing human breast cancer cells destruction using combination of adenovirus expressing P53 and hyperthermia treatment

In Malaysia, breast cancer is the most common cancer where 1 in 19 Malaysian women will be diagnosed with breast cancer by the age of 85. Moreover, lack of specific symptoms in the early stage of disease leading to delay in diagnosis. Unfortunately, current treatments by chemotherapeutic agents, surgery and radiation are not fully effective for the treatment of breast cancer. Thus, there is an urgency in developing new approaches for the treatment of breast cancer patients. In this study, a novel therapeutic regimen, combining the effects of recombinant adenovirus and hyperthermia was investigated. Firstly, Adenovirus serotype 5 was constructed by cloning of p53 gene into a defective recombinant adenovirus vector, Ad5-p53-DsRed Monomer N1. The Ad5-p53-DsRed Monomer N1 (MOI of 100) was then used to infect breast cancer cells (MDA-MB 231 and MCF-7) with or without combination of hyperthermia treatment (42ºC for 2 hours). The cell killing and viral concentration were then determined by MTT assay and viral plaque formation assay respectively. After that, the heat shock protein (Hsp70) and p53 protein expression in transfected cells were quantitated using ELISA assay. Activated-Caspase 3/7, 8 and 9 were also evaluated to study the apoptotic pathway of cancer cells. Furthermore, the novel protein interaction between nucleotide binding domain (NBD) Hsp70 and human Ad5 E1A 32 kDa motif (PNLVP); and NBD and p53 motif (SCMGGMNR) were investigated through bioinformatics tools such as Gromacs and Autodock softwares. It was found that MDA-MB 231 and MCF-7 cells infected with virus Ad5-p53- DsRed Monomer N1 alone resulted in 46.77±2.74% and 42.26±1.78% cell killing respectively while hyperthermia in combination with virus were 84.82±1.64% and 80.13±3.30% respectively. The Hsp70 expression of both cancer cells was also increased to 170.57% (MDA-MB 231) and 169.83% (MCF-7). Moreover, p53 expression in MDA-MB 231 and MCF-7 cells by virus combined with heat treatment (85.72 ng/L and 79.05 ng/L respectively) could lead to enhanced oncolytic property compared to virus treatment alone (47.82 ng/L and 40.54 ng/L respectively). In addition, caspase activity was first time reported that apoptosis process started at very early stage of infection in breast cancer cells with hyperthermia compared to virus alone. This was due to the evident that the highest kinetic energy was found in caspase 3 whereas virus alone the highest in caspase 8. In conclusion, Hsp70 induction by hyperthermia treatment enhanced Ad5-p53-DsRed Monomer N1 replication and oncolysis in MDA-MB 231 and MCF-7 cells through apoptotic pathway. Besides that, NBD of Hsp70 had the best interaction with PNLVP motif at 42°C. Thus, combining Ad5-p53 with hyperthermia treatment could be a potential approach for breast cancer treatment

Antitumor effect of fever range whole body hyperthermia with curcumin in breast cancer-induced mice

Breast cancer is a complex and heterogeneous disease and also one of the major cancer types among female worldwide. Fever range whole-body hyperthermia and curcumin as a single treatment have been tested against breast cancer and showed some promising anti-tumor effect. However, their combination as an effective anti-tumor treatment against breast cancer has never been explored. The effects of combined whole-body fever range hyperthermia and curcumin on tumor growth was examined in this study. Mice were inoculated with EMT6 cells subcutaneously and allocated to 4 treatment groups: (i) control (control), (ii) curcumin (50mg/kg bodyweight), (iii) twice fever range whole-body hyperthermia 39.0°C (± 0.5) for 15 minutes, (iv) a combination of curcumin (50mg/kg bodyweight) and twice fever range whole-body hyperthermia 39.0°C (± 0.5) for 15 minutes. Following treatment, mice body weight and tumor volume were measured. The greatest tumor growth inhibition exhibited in combination treatment (68.45%, p<0.05) and showed no general toxicity. As conclusion, the combination treatment can be a potential anti-tumor treatment of breast cancer

Hyperthermia combined with chemotherapy - Biological rationale, clinical application, and treatment results

There is substantial evidence from preclinical data that the antitumor cytotoxicity of selected chemotherapeutic agents either alone or combined with radiation can be enhanced by appropriate heat exposure (40-44 degrees C) of cells or tumor tissues. Based upon these results the integration of hyperthermia as an additional treatment modality, given simultaneously with systemic chemotherapy or in combination with radiochemotherapy, is currently tested at the clinic. Regional hyperthermia combined with chemotherapy or radiochemotherapy showed impressive results (phase II studies) at clinical relevant temperatures in locally advanced tumors of different entities in terms of objective response rate, local tumor control and relapse-free survival. Clinical protocols of well-designed phase III trials on combined treatment modalities integrating hyperthermia are rather limited but for some tumors confirm its clinical benefit. In general, the clinical approach to use hyperthermia has gained much more interest within in the field of medical oncology. One of the major reason is the substantial technical improvements made with the available commercial equipment for local or regional heating, especially in case of deep-seated lesions or systemic heating. Further testing of the potential of hyperthermia combined with chemotherapy or radiochemotherapy in prospective randomized trials are warranted. At this time, hyperthermia as an adjunct to conventional treatment strategies is recommended in the setting of clinical protocols. The results of prospective trials should answer the question for which types of local advanced or metastatic tumors hyperthermia becomes standard as part of a multi-modal treatment strategy

Focal Spot, Fall/Winter 1986

https://digitalcommons.wustl.edu/focal_spot_archives/1044/thumbnail.jp

Focal Spot, Spring 1987

https://digitalcommons.wustl.edu/focal_spot_archives/1045/thumbnail.jp

Focal Spot, Summer 1986

https://digitalcommons.wustl.edu/focal_spot_archives/1043/thumbnail.jp

Locoregional hyperthermia of deep-seated tumours applied with capacitive and radiative systems. A simulation study

Background: Locoregional hyperthermia is applied to deep-seated tumours in the pelvic region. Two very different heating techniques are often applied: capacitive and radiative heating. In this paper, numerical simulations are applied to compare the performance of both techniques in heating of deep-seated tumours. Methods: Phantom simulations were performed for small (30 × 20 × 50 cm 3 ) and large (45 × 30 × 50 cm 3 ), homogeneous fatless and inhomogeneous fat-muscle, tissue-equivalent phantoms with a central or eccentric target region. Radiative heating was simulated with the 70 MHz AMC-4 system and capacitive heating was simulated at 13.56 MHz. Simulations were performed for small fatless, small (i.e. fat layer typically 3 cm) patients with cervix, prostate, bladder and rectum cancer. Temperature distributions were simulated using constant hyperthermic-level perfusion values with tissue constraints of 44 °C and compared for both heating techniques. Results: For the small homogeneous phantom, similar target heating was predicted with radiative and capacitive heating. For the large homogeneous phantom, most effective target heating was predicted with capacitive heating. For inhomogeneous phantoms, hot spots in the fat layer limit adequate capacitive heating, and simulated target temperatures with radiative heating were 2–4 °C higher. Patient simulations predicted therapeutic target temperatures with capacitive heating for fatless patients, but radiative heating was more robust for all tumour sites and patient sizes, yielding target temperatures 1–3 °C higher than those predicted for capacitive heating. Conclusion: Generally, radiative locoregional heating yields more favourable simulated temperature distributions for deep-seated pelvic tumours, compared with capacitive heating. Therapeutic temperatures are predicted for capacitive heating in patients with (almost) no fat

Thermal dosimetry for bladder hyperthermia treatment. An overview.

The urinary bladder is a fluid-filled organ. This makes, on the one hand, the internal surface of the bladder wall relatively easy to heat and ensures in most cases a relatively homogeneous temperature distribution; on the other hand the variable volume, organ motion, and moving fluid cause artefacts for most non-invasive thermometry methods, and require additional efforts in planning accurate thermal treatment of bladder cancer. We give an overview of the thermometry methods currently used and investigated for hyperthermia treatments of bladder cancer, and discuss their advantages and disadvantages within the context of the specific disease (muscle-invasive or non-muscle-invasive bladder cancer) and the heating technique used. The role of treatment simulation to determine the thermal dose delivered is also discussed. Generally speaking, invasive measurement methods are more accurate than non-invasive methods, but provide more limited spatial information; therefore, a combination of both is desirable, preferably supplemented by simulations. Current efforts at research and clinical centres continue to improve non-invasive thermometry methods and the reliability of treatment planning and control software. Due to the challenges in measuring temperature across the non-stationary bladder wall and surrounding tissues, more research is needed to increase our knowledge about the penetration depth and typical heating pattern of the various hyperthermia devices, in order to further improve treatments. The ability to better determine the delivered thermal dose will enable clinicians to investigate the optimal treatment parameters, and consequentially, to give better controlled, thus even more reliable and effective, thermal treatments