Photodiagnosis and Photodynamic Therapy (2015) 12, 146—149
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SHORT COMMUNICATION
Study of low doses cisplatin synergistic effect on photodynamic outcome of aluminum phythalocyanine on soft tissue sarcoma (RD) cell line Safdar Ali Ph.D. a,b,∗, Ahmat Khurshid b, M. Maqsood b, M. Rafi b, Junaid A. Khan b, S.S.Z. Zaidi c, Saleh Mohammad a, Masroor Ikram b a
Department of Physics, Hazara University, Mansehra, Pakistan Department of Physics and Applied Mathematics, Pakistan Institute of Engineering and Applied Sciences, Islamabad 45650, Pakistan c National Institute of Health, Chawk Shehzad, Park Road, Islamabad, Pakistan Available online 3 January 2015 b
KEYWORDS Photodynamic therapy; Cisplatin; Rhabdomyosarcoma; Neutral red; Cell viability; Diode laser
Summary Photodynamic therapy (PDT) in combination with other treatment modality expects to overcome the drug resistance experienced in monotherapy. In this present work combination of chemo cum PDT is studied over the range of doses. It is found that treating cells/exposing cells to chemo drug (cisplatin, CDDP) and PDT individually results in minimal cell killing (∼7% and ∼16%) compared to the administration of chemo followed by PDT (∼50% cells were viable). These results showed that cell viability synergistically decreases in case of combination treatment as compared with individual treatment. Photodynamic therapy (PDT) in combination with CDDP chemotherapy expects to overcome the drug resistance experienced in monotherapy. © 2015 Elsevier B.V. All rights reserved.
Introduction Soft tissue sarcoma is the major cause of cancer in children and is a type that usually develops in voluntary muscles.
∗ Corresponding author at: Hazara University, Physics, Khyber Pukhtoonkhwa, Dhodial, Mansehra 21300, Pakistan. Tel.: +092 997 414132; fax: +092 997 530046. E-mail addresses:
[email protected],
[email protected] (S. Ali).
http://dx.doi.org/10.1016/j.pdpdt.2014.12.011 1572-1000/© 2015 Elsevier B.V. All rights reserved.
It has also potentential to grow anywhere in a body irrespective of voluntary muscles cells but more favorable site is head and neck, abdomen or one of extremities [1—3]. Children who develop metastatic rhabdomyosarcoma have very poor prognosis. About 65% cases are diagnosed at the age of 6 or below while the remaining may occur up to the age of 18 [4,5]. Unfortunately, children with metastatic RMS disease prognosis have not improved significantly [2,6—8]. Beside the side effects and treatment cost of late stages, most established treatment modalities are radio-, chemo-, immunotherapy, surgery etc., against soft tissue sarcoma
Low doses cisplatin synergistic effect on soft tissue sarcoma
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[9,10]. Photodynamic therapy is relatively new less invasive and emerging oncological and non-oncological treatment modality [11—14]. Researchers are focusing in exploring low doses regimes both for photodynamic drug and chemo in-order to avoid complications [15,16]. In this present study cisplatin also known as CDDP been used in combination with photodynamic therapy to enhance their accumulative therapeutic outcome. Combinational therapy (CDDP cum PDT) been reported for various cancers i.e., lungs, cervical etc., in a number of published research works [17—20].
Materials and methods Cell lines and chemicals Rhabdomysarcoma cell (RD) line was provided by National Institute of Health (NIH) Islamabad. Cell culture media RPMI-1640 with 10% FBS (Invitrogen) was used. Cisplatin (CDDP) was provided by Nuclear Medicine Oncology and Radiotherapy Institute (NORI) Pakistan. Aluminum pathalocyanin tetrasulphonated chloride (AlPs-4, photosense) was manufactured by Russian Pharmaceutical Company. Stock solution concentration of 3.33 mM of both CDDP and aluminum phthalocyanine tetrasulfonate chloride (AlS4 Pc-Cl) were stored at −20 ◦ C. Neutral red (NR) was purchased from Sigma Aldrich for cellular viability.
Photo- and dark-toxicity RD cells were cultured in flat bottom 96-well plate. Post 24 h of seeding one set of culture was exposed to diode laser of 630 ± 5 nm (red) delivered energy doses in the range of doses (5—55 J/cm2 while another set of culture was exposed to range of concentrations of photosensitizer (PS) with incubation time of 45 min. Neutral red (NR) assay was performed for the cellular viability calculation.
Fig. 1 % Cell viability with (a) various dose of light (5—55 J/cm2 ) and (b) various concentrations (5—30 M) of photosensitizer.
Chemotherapy and photodynamic treatment Post confluent cell culture grown in 96-well plate, chemo drug with different concentrations administered for 24 h. For photodynamic therapy, at the end of PS incubation time culture was irradiated with 10 J/cm2 of laser.
Combinational treatment Cell culture was incubated with different concentrations of cisplatin for 24 h followed by 10 M PS for 45 min. Photodynamic therapy (10 J/cm2 ) was carried out in the presence of chemo-drug. To evaluate the synergistic effects of chemoPDT treatment NR-assay was performed.
Results Present work is focused on to minimize the dose of cytotoxic drugs in combination of PDT. Anti cancer drugstarget
several cellular components and activate responses ranging from cell repair to cell death. From Fig. 1a and b non-toxic region for both light (10 J/cm2 ) and PS (10 M) individually was selected for further study. Varying concentration of PS was selected for PDT step while light dose was kept constant. It was observed that cellular viability is dose dependent such as decreases with increasing concentration (Fig. 2a). In case of chemotherapy with CDDP the viability decreases as the concentration of CDDP increases. In the range of 5—25 M the viability decreases from 93% to 73% (Fig. 2b). Chemo drug mode of action is cross linking through inter-strand and/or intra-strand of DNA. The lower end of incubated concentration has more viable cells depicting also fewer side effects related to dose dependency to normal tissues. Low doses of chemo drug results in cells growth arrest which offer favorable participation for combination therapy, aiming to have synergistic effect of PDT and chemo low doses.
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Conclusions Chemotherapy being first choice of treatment is effective only at high doses, because at low doses tumor cells has tendency to repair initiated damages thereby possibly resume their original high proliferation rate. Whereas in case of low doses of PS in PDT arrest cells in G2—M phase leading to apoptosis, cytostatic condition [21,22]. CDDP inhibits DNA replication and chain elongation, which accounts for its antineoplastic activity [23]. Fig. 3 result of low dose CDDP and PDT in combination have resemblance with that of the previous studies conducted on combination therapy [16,17]. Both CDDP and PDT in combination arrest cells growth and division in S—G2 and G2 —M phase, respectively [21,24,25]. Our results show a clear anti-cancer activity of low doses of PDT and CDDP in combination on rhabdomyosarcoma cell line [19—22]. Our results give a new pathway for treating cancer to minimize anti-cancer agent dose side effects. Therefore, our study suggested that low doses of photosensPDT in combination with low doses CDDP is an effective therapy for rhabdomyosarcoma.
Acknowledgements We acknowledge the Higher Education Commission (HEC), Pakistan for the financial support through the Indigenous Fellowship Scheme for PhD Studies in Science & Technology.
References
Fig. 2 % Viability of cell with (a) various concentration of PS at constant light dose and (b) chemo-drug (5—55).
Lower arm of both the PS-PDT and Chemo was selected and proceed for combinational therapy. In combination therapy low dose of CDDP (10 M) and PDT (10 M, 10 J/cm2 ) have additive effect Fig. 3.
Fig. 3 Comparison of different treatment arms CDDP, PDT and CDDP + PDT effect on cell viability.
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