Background: Despite its anticancer activity, cisplatin exhibits severe testicular toxicity when used in chemotherapy. Owing to its wide application in cancer therapy, the reduction of damage to normal tissue is of imminent clinical need. In this study, we evaluated the effects of catechin hydrate, a natural flavon-3-ol phytochemical, on cisplatin-induced testicular injury. Methods: Type 2 mouse spermatogonia (GC-1 spg cells) were treated with 0-100 μM catechin and cisplatin. Cell survival was estimated using a cell proliferation assay and Ki-67 immunostaining. Apoptosis was assessed via flow cytometry with the Dead Cell Apoptosis assay. To determine the antioxidant effects of catechin hydrate, Nrf2 expression was measured using qPCR and CellROX staining. The anti-inflammatory effects were evaluated by analyzing the gene and protein expression levels of iNOS and COX2 using qPCR and immunoblotting. Results: The 100 μM catechin hydrate treatment did not affect healthy GC-1 spg cells but, prevented cisplatin-induced GC-1 spg cell death via the regulation of anti-oxidants and inflammation-related molecules. In addition, the number of apoptotic cells, cleaved-caspase 3 level, and BAX gene expression levels were significantly reduced by catechin hydrate treatment in a cisplatin-induced GC-1 spg cell death model. In addition, antioxidant and anti-inflammatory marker genes, including Nrf2 , iNOS, and COX2 were significantly downregulated by catechin hydrate treatment in cisplatintreated GC-1 cells. Conclusions: Our study contributes to the opportunity to reintroduce cisplatin into systemic anticancer treatment, with reduced testicular toxicity and restored fertility.

Background: Brassica oleracea var. italica (broccoli), a rich source of antioxidants, can prevent various diseases and improve human health. In this study, we investigated the antioxidative effects of broccoli sprout extract on oxidative stress induced by lipopolysaccharide and cisplatin in cell and organ tissue models. Methods: Antioxidative effect of BSE was evaluated using DPPH and ABTS in RAW 364.7 cells, and effects of BSE on testes were investigated using Cisplatin-induced testicular damage model with an in vitro organ culture system. Results: The DPPH assay showed that the antioxidant activity of the alcoholic broccoli sprout extract was higher than that of the water extract. Additionally, the expression levels of antioxidation-related genes, Nrf2 , Gsr , HO-1, and catalase , were significantly increased in broccoli sprout extract-treated RAW 264.7 cells, and the extract suppressed lipopolysaccharide-induced mitochondrial dysfunction. Based on the results in the RAW 264.7 cell culture, the antioxidative effects of the extracts were investigated in a mouse testis fragment culture. The expression of Nrf2 , HO-1 , and Ddx4 was clearly decreased in cisplatin-treated mouse testis fragments and not in both broccoli sprout extract- and cisplatin-treated mouse testis fragments. In addition, the oxidative marker O-HdG was strongly detected in cisplatin-treated mouse testis fragments, and these signals were reduced by broccoli sprout extract treatment. Conclusions: The results of this study show that broccoli sprout extracts could serve as potential nutraceutical agents as they possess antioxidant effects in the testes.

시스플라틴은 지난 몇 십년간 가장 많이 사용되고 있는 항암제 중 하나이다. 2가의 시스플라틴의 가장 큰 단점은 암세포 뿐만 아니라 정상세포도 공격하여 부작용을 나타낸다. 본 연구에서는 이러한 단점을 줄이기 위해 목표로 한 타겟을 공격하여 빠른 시간 내에 암세포를 죽일 수 있는 4가의 백금화합물 3과 4를 합성하였다. 합성된 Pt(IV)-Bu2 3 and Pt(IV)-Val2 4는 NMR과 질량분석기를 통하여 구조를 확인 하였다. 합성된 Pt(IV) 화합물 3과 4는 위암세포주인 MCF-7을 대상으로 한 MTT assay를 통해 항암효과를 조사하였다. 그 결과 시스플라틴은 39%, Pt(IV)-Bu2 3와 Pt(IV-Val2 4는 50μM 농도에서 각각 54%와 84%의 세포사멸을 보였다.

Background: Cisplatin is a well-known platinum-containing anti-cancer drug against bladder, ovarian, lung and testicular cancer. However, the potential effects and molecular targets of cisplatin in human mucoepidermoid carcinoma (MEC) are not fully understood. Here, we investigated the apoptotic effect and underlying mechanism of cisplatin in human MEC cells. Methods: The potential effects of cisplatin were evaluated by trypan blue exclusion assay, Western blotting, 4’-6-diamidino-2-phenylindole (DAPI) staining, live/dead assay and immunocytochemistry. Results: Cisplatin suppressed cell growth and enhanced expression of cleaved PAPR in MC3 and YD15 cells. Cisplatin caused morphological change of nuclei and increased the number of ethidium homodimer-1-stained cells. In addition, cisplatin commonly increased Bax activation in both cells, while other Bcl-2 family proteins were not affected. Conclusions: These results suggest that cisplatin might induce apoptosis by activating Bax protein, which would provide baseline data for development of effective treatment strategy against MEC.

Immunoprecipitation-based high performance liquid chromatography (IP-HPLC) is a type of modified enzyme-linked immunosorbent assay (ELISA) that uses protein A/G (or antibody)-conjugated beads instead of the antibody-conjugated wells used in ELISA. In order to determine the fidelity of IP-HPLC, the author used 83 antisera to identify protein expression changes caused by cisplatin treatment in KB human oral cancer cells. KB cells were cultured for 12 or 24 hours with 10 ug/mL cisplatin. The results obtained by IP-HPLC were comparable with published cisplatin data, although ELISA was not conducted in the present study. Cisplatin dominantly reduced the levels of proteins associated with cell proliferation, transcription factors, growth factors, cytoskeletal proteins, and cellular differentiating factors, but on the other hand, apoptosis-related factors, oncogenes, and protective proteins were usually up-regulated, presumably to address cisplatin-induced DNA damage. In particular, cisplatin directly inactivated genomic DNA by down-regulating histone H1 and demethylase and by up-regulating deacetylase. Cisplatin also rapidly induced p53 overexpression and mitochondria-mediated endogenous apoptosis occurred after 12 hours of cisplatin treatment, although this was almost completely replaced by FASL/FAS-mediated exogenous apoptosis after 24 hours. This preliminary study was conducted to investigate the anticancer effect of cisplatin on the KB human oral cancer cells and to determine the fidelity of IP-HPLC data. It was concluded that IP-HPLC is useful for identifying profile changes of genome wide essential proteins and signaling changes of major molecular pathways.

Curcumin is a widely used flavoring agent in food, and it has been reported to inhibit cell growth, to induce apoptosis, and to have antitumor activity in many cancers. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatment with curcumin and cisplatin on human tongue SCC25 cells. To investigate whether the co-treatment efficiently reduced the viability of the SCC25 cells compared with the two treatments separately, an MTT assay was conducted. The induction and the augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and an analysis of DNA hypoploidy. Western blot, MMP and immunofluorescence tests were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following the co-treatment. In this study, following the co-treatment with curcumin and cisplatin, the SCC25 cells showed several forms of apoptotic manifestation, such as nuclear condensation, DNA fragmentation, reduction of MMP, increased levels of Bax, decreased levels of Bcl-2, and decreased DNA content. In addition, they showed a release of cytochrome c into the cytosol, translocation of AIF and DFF40 (CAD) to the nuclei, and activation of caspase-7, caspase-3, PARP, and DFF45 (ICAD). In contrast, separate treatments of 5 μM of curcumin or 4 μg/ml of cisplatin, for 24 hours, did not induce apoptosis. Therefore, our data suggest that combination therapy with curcumin and cisplatin could be considered as a novel therapeutic strategy for human oral squamous cell carcinoma.

Eugenol (4-allyl-2-methoxyphenol) is a naturally occurring phenolic compound that is widely used in dentistry as a component of zinc oxide eugenol cement that is commonly applied to the mouth environment. Cisplatin is one of the most potent known anticancer agents and shows significant clinical activity against a variety of solid tumors. This study was undertaken to investigate the synergistic apoptotic effects of co-treatments with eugenol and cisplatin on human melanoma (G361) cells. To investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment, an MTT assay was conducted. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining and an analysis of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate the expression levels and the translocation of apoptosis-related proteins following this co-treatment. Furthermore, proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed. The results indicated that a co-treatment with eugenol and cisplatin induced multiple pathways and processes associated with an apoptotic response in G361 cells including nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, the increase and decrease of Bax and Bcl-2, a decreased DNA content, the release of cytochrome c into the cytosol, the translocation of AIF and DFF40 (CAD) into the nucleus, and the activation of caspase-9, caspase-7, caspase-3, PARP and DFF45 (ICAD). In contrast, separate treatments of 300 μM eugenol or 3 μM cisplatin for 24 h did not induce apoptosis. Our present data thus suggest that a combination therapy of eugenol and cisplatin is a potential treatment strategy for human melanoma.

Photodynamic therapy(PDT) is recently developed as an effective treatment for malignant disease. Carboplatin, a less nephrotoxic analog of cisplatin, has been widely used for the treatment of multiple malignancies. In this study, we investigated the cytotoxic and apoptotic effect of combined modality of photofrin mediated PDT with cisplatin and carboplatin on KB cell human oral cancer cell line in vitro. The a ttached KB cells were incu bated with c isplatin(0.04mg/ml) and carboplatin(0.02mg/ml) for 24h at 37℃ and followed by photosensitization with photofrin for 6h and laser irradiation with 630nm LED at an intensity of 2.0 J/cm2 for activating photofrin for 15min. Then MTT assay and SYTO 16 green & Propidium iodide (PI) double staining were used respectively to measure the cytotoxicity and nuclear morphology at 24h after PDT. This study demonstrates that the combined modality with carbopaltin resulted in enhanced apoptotic cell death as well as cytotoxic e ffect on KB c ells in vitro, which s uggests the feasibility of combined modality and the possibility o f reducing the effective dosage of photofrin and carboplatin and lowering the side effects on normal cells

Cisplatin(ctr-diamminedichlorop1atium) is one of the most effective anti-cancer drugs being clinically used in the treatment of solid tumors. Despite its therapeutic benefits, its use in clinical practice is often limited because of dose related toxicity. It is known that yeast cell wall beta-glucans possess immuno-modulating properties, which allows for their application in antitumor therapy. 1S2 is a kind of beta-glucan derived from the cell wall of mutated Sacchammyces cerevisiae, which exhibits anti-cancer activity in vitro and in vivo. The present study explored the possibility of combination therapy of IS2 with cisplatin. In experimental metastasis of colon26M3.1 cells, prophylactic intravenous administration of IS-2 in combination with cisplatin effectively inhibited tumor metastasis compared with cisplatin alone or IS-2 treatment in vivo. IS-2 effectively enhanced Thl type cytokines including IFN-r, IL-2, ILf2 and GM-CSF. Simultaneously, this combined treatment inhibited production of Th2 type cytokines compared with control. These results suggested that IS-2 can be applied in combination therapy with anti-cancer drugs to minimize their side effects.

Gemcitabine (Gemzar, 2,2-difluorodeoxycytidine, or dFdC) is an analog of cytosine arabinoside with anti-tumor activity in a few human cancers (lung, ovary, pancreatic and breast cancers). However, the mechanism of apoptosis by this compound in carcinoma has not been fully elucidated. In the present study, we investigated that gemcitabine alone and combination with cisplatin or 5-FU are cancer toxicity using lung cancer cell line A549 by MTT, FACS analysis, and Western blot assay. Also, to confirm enhanced antitumor activity in vivo using an xenograft tumor model. The MTT assay showed higher cytotoxic effect in combination with gemcitabine-cisplatin or gemcitabine-5-FU than gemcitabine alone. FACS analysis showed that gemcitabine-cisplatin combination increased hypodiploid DNA to 70.84 %. The induction of apoptosis showed more increase in combination with gemcitabine-cisplatin or gemcitabine-5-FU than gemcitabine alone. The Western results showed higher expression of p53 and p21WAF/CIP1 protein in combination treatment with gemcitabine-cisplatin or gemcitabine-5-FU than gemcitabine treatment alone. But, Bcl-2 protein expression decreased. In vivo experiments showed that more decreased tumor size and more increased survival rate on combination with gemcitabine- cisplatin or gemcitabine-5-FU combination than gemcitabine alone. In conclusion, this study suggests that gemcitabine combined with cisplatin or 5-FU are the synergistic effect of anticancer therapy on lung cancer.

Background: Cisplatin is one of the most extensively used chemotherapeutic agents for the treatment of cancer, including bladder, and ovarian cancers. However, it has been shown to induce nephrotoxicity, despite being an outstanding anti-cancer drug. In this study, we investigated the protective effect of dopaol β-D-glucoside (dopaol) on cisplatin-induced nephrotoxicity. Methods and Results: To confirm the protective effect of dopaol on cisplatin-induced nephrotoxicity, HK-2 cells were treated with 20 μM cisplatin and 80 μM dopaol. Cisplatin increased apoptosis, caspase-3 activity and mitochondrial dysfunction; however pretreatment with 80 μM dopaol successfully attenuated apoptosis, caspase-3 activity and mitochondrial dysfunction. To evaluate the protective effect dopaol on cisplatin-induced nephrotoxicity in vivo, we used an animal model (balb/c mice, 20 ㎎/㎏, i.p. once/day for 3 day). The results were similar to those obtained using HK-2 cells; renal tubular damage and neutrophilia induced by cisplatin reduced following dopaol injection (10 ㎎/㎏, i.p. once/day for 3 day). Conclusions: These results indicate that dopaol treatment reduced cisplatin-induced nephrotoxicity in vitro and in vivo, and can be used to treat cisplatin-induced nephrotoxicity. However, further studies are required to determine the toxicity high dose dopaol and the signal pathways involved in its mechanism of action in animal models.

Background : In this study, we investigated the renoprotective effects of serotonin and its derivatives, on the renal function and expression of inflammation and apoptosis in cisplatin-induced nephrotoxicity mice. Methods and Results : Serotonin and its derivatives were orally administered at a dose of 7.5 mg/kg body weight for 5 days before the intraperitoneal injection of cisplatin 20 mg/kg body weight, and the effects were compared with those of vehicle-treated nephrotoxicity control and normal groups. In the serum and kidney, renal function parameters, reactive oxygen species and expression of protein related to pro-oxidant, antioxidant, inflammation and apoptosis were examined. As a result, serotonin and its derivatives administrations to nephrotoxicity mice lowered serum BUN and creatinine concentrations. These results were derived, at least in part, from attenuation the expression of antioxidant enzymes-related proteins, SOD and GPx. In the cisplatin-induced renal condition, augmented p-p38, p-ERK and p-JNK (mitogen-activated protein kinase pathway) were reduced with a increase in antioxidant enzymes on serotonin and its derivatives treatment. Moreover, in the serotonin and its derivatives-treated groups, NF- κB-induced inflammatory factors and apoptotic protein expressions were regulated in the kidney. Conclusion : The present study indicates that serotonin and its derivatives exerts a renoprotective effect against cisplatin-induced nephrotoxicity through the recovery of kidney function deterioration and attenuation of renal inflammation and apoptosis by regulating oxidative stress condition.