• Asian Pacific Journal of Cancer Prevention
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• v.15 no.2
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• pp.617-622
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• 2014

Voacanga globosa (Blanco), a plant endemic to the Philippines, is traditionally used especially by indigenous people of Bataan in the treatment of ulcers, wounds and tumorous growths. This study aimed to provide scientific evidence to therapeutic properties by determining cytotoxic and pro-apoptotic activity of HPLC fractions from leaves on HCT116 human colon carcinoma and A549 human lung carcinoma cell lines. Ethanolic extraction was performed on V globosa leaves followed by hexane and ethyl acetate partitioning. Silica gel column chromatography and high performance liquid chromatography (HPLC) produced MP1, MP2 and MP3 fractions. Cytotoxic activity of the fractions was determined through MTT assay against the cancer cell lines HCT116 and A549 and the non-cancer AA8 Chinese hamster ovarian cell line. Pro-apoptotic activities of the most active fractions were further assessed through DAPI staining, TUNEL assay and JC-1 mitochondrial membrane potential assay with HCT116 cells. While the MPI fraction exerted no significant activity against all cell lines tested, MP2 and MP3 fractions demonstrated high toxicity against HCT116 and A549 cells. The MP3 fraction induced formation of apoptotic bodies, condensed DNA and other morphological changes consistent with apoptosis of HCT116 cells and TUNEL assay showed significant increase in DNA fragmentation over time. In these cells, the MP3 fraction also induced mitochondrial membrane destabilization, which is generally associated with the beginning of apoptosis. Phytochemical analysis demonstrated the presence only of saponins and terpenoids in the MP3 fraction. The results indicate that the MP3 fraction exerts cytotoxic activity on HCT116 cells via induction of apoptosis triggered by loss of mitochondrial membrane potential crucial for cell survival.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.5
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• pp.3191-3196
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• 2013

Leaf extracts of Cassia alata L (akapulko), traditionally used for treatment of a variety of diseases, were evaluated for their potential antitumor properties in vitro. MTT assays were used to examine the cytotoxic effects of crude extracts on five human cancer cell lines, namely MCF-7, derived from a breast carcinoma, SK-BR-3, another breast carcinoma, T24 a bladder carcinoma, Col 2, a colorectal carcinoma, and A549, a nonsmall cell lung adenocarcinoma. Hexane extracts showed remarkable cytotoxicity against MCF-7, T24, and Col 2 in a dose-dependent manner. This observation was confirmed by morphological investigation using light microscopy. Further bioassay-directed fractionation of the cytotoxic extract led to the isolation of a TLC-pure isolate labeled as f6l. Isolate f6l was further evaluated using MTT assay and morphological and biochemical investigations, which likewise showed selectivity to MCF-7, T24, and Col 2 cells with $IC_{50}$ values of 16, 17, and 17 ${\mu}g/ml$, respectively. Isolate f6l, however, showed no cytotoxicity towards the non-cancer Chinese hamster ovarian cell line (CHO-AA8). Cytochemical investigation using DAPI staining and biochemical investigation using terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-a method used to detect DNA fragmentation-together with caspase assay, demonstrated apoptotic cell death. Spectral characterization of isolate f6l revealed that it contained polyunsaturated fatty acid esters. Considering the cytotoxicity profile and its mode of action, f6l might represent a new promising compound with potential for development as an anticancer drug with low or no toxicity to non-cancer cells used in this study.

• Journal of Physiology & Pathology in Korean Medicine
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• v.19 no.1
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• pp.224-229
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• 2005

Ginseng radix, the root of Panax ginseng C.A.Meyer (Araliaceae), has traditionally been used for the treatment of various disorders including cerebrovascular accident (CVA). In the present study, the effect of Ginseng radix on c-Fos expression and apoptosis in the hippocampal CA1 region of gerbils following transient global ischemia was investigated via immunohistochemistry for c-Fos and caspase-3 and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. Enhanced c-Fos-, TUNEL-, and caspase-3-positivities were detected in the hippocampal CA1 region in ischemic gerbils. Administration of the aqueous extract of Ginseng radix suppressed this ischemia-induced increment in the numbers of c-Fos-, TUNEL-, and caspase-3-positive cells. These results suggest that Ginseng radix has an inhibitive effect on the induction of c-Fos expression and apoptosis seen following transient global ischemia.

• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.233-233
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• 2004

PURPOSE: Gene expression and apoptosis in testicular germ cells has been demonstrated in many transgenic animals. However, little is known about the transgenic pig and rates of apoptosis during spermatogenesis. METHODS : Morphological and biochemical features of apoptosis reported in other species were used to confirm that the TdT-mediated dUTP Nick end labeling (TUNEL) assay is an acceptable mothos for idendtification and quantification of apoptotic transgenic germ cells in histological tissue section from transgenic pig testis. (omitted)

• International Journal of Oral Biology
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• v.36 no.1
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• pp.31-35
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• 2011

Teeth develop via a reciprocal induction between the ectomesenchyme originating from the neural crest and the ectodermal epithelium. During complete formation of the tooth morphology and structure, many cells proliferate, differentiate, and can be replaced with other structures. Apoptosis is a type of genetically-controlled cell death and a biological process arising at the cellular level during development. To determine if apoptosis is an effective mechanism for eliminating cells during tooth development, this process was examined in the rat mandible including the developing molar teeth using the transferase-mediated dUTP-biotin nick labeling (TUNEL) method. The tooth germ of the mandibular first molar in the postnatal rat showed a variety of morphological appearances from the bell stage to the crown stage. Strong TUNEL-positive reactivity was observed in the ameloblasts and cells of the stellate reticulum. Odontoblasts near the prospective cusp area also showed a TUNEL positive reaction and several cells in the dental papilla, which are the forming pulp, were also stained intensively in this assay. Our results thus show that apoptosis may take place not only in epithelial-derived dental organs but also in the mesenchyme-derived dental papilla. Hence, apoptosis may be an essential biological process in tooth development.

• Korean Journal of Pharmacognosy
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• v.36 no.2 s.141
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• pp.88-92
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• 2005

Apoptosis plays an important role in autoimmune chronic (Hashimoto's) thyroiditis, a disorder that often results in hypothyroidism. The goal of this study was to induce apoptosis by the combination of inflammatory cytokines, interferon $(IFN)-{\gamma}$ and tumor necrosis factor $(TNF)-{\alpha}$, and to investigate a potential role of medicinal plants in the thyroid follicular cells (FRTL) in vitro. The apoptosis was evaluated by cellular viability, DNA fragmentation, and terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) assay. Extract of Gamgung-tang (GGT, Glycyrrhizae Radix, black beans, Angelicae Radix, and Cnidii Rhizoma) $(0.3{\sim}9.0mg/ml)$ was shown to maintain the viability of cells treated with $IFN-{\gamma}(100U/ml)$ and $TNF-{\alpha}$ (0.5 ng/ml). FRTL cells were found to undergo DNA fragmentation with the inflammatory cytokines. The extract of GGT inhibited DNA fragmentation in dose-dependent manner. The cells with TUNEL-positive nuclei were detected with $IFN-{\gamma}$ and $TNF-{\alpha}$ treatment. The number of TUNEL-positive cells decreased with the treatment of extract of GGT. These results indicate that medicinal plants inhibit the occurrence of apoptosis in thyroid follicular cells, therefore, may have therapeutic potential in the treatment of autoimmune chronic thyroiditis.

• Nutrition Research and Practice
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• v.3 no.3
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• pp.180-184
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• 2009

The apoptotic effect of bacteria-derived $\beta$-glucan was investigated in human colon cancer cells SNU-C4 using terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, reverse transcription-polymerase chain reaction (RT-PCR) expressions of Bcl-2, Bax, and Caspase-3 genes, and assay of caspase-3 enzyme activity. $\beta$-Glucan of 10, 50, and $100{\mu}g$/mL decreased cell viability in a dose-dependent manner with typical apoptotic characteristics, such as morphological changes of chromatin condensation and apoptotic body formation from TUNEL assay. In addition, $\beta$-glucan ($100{\mu}g$/mL) decreased the expression of Bc1-2 by 0.6 times, whereas the expression of Bax and Caspase-3 were increased by 3.1 and 2.3 times, respectively, compared to untreated control group. Furthermore, the caspase-3 activity in the $\beta$-glucan-treated group was significantly increased compared to those in control group (P < 0.05). Bacterial derived $\beta$-glucan could be used as an effective compound inducing apoptosis in human colon cancer.

• Journal of Acupuncture Research
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• v.24 no.4
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• pp.107-113
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• 2007

목적 : 본 연구는 계혈등 약침액이 자궁경부암 세포주 SNU-17에서 세포 사멸 효과가 있는지 알아보고자 하였다. 방법 : 자궁경부암 세포주 SNU-17에서 세포 사멸의 변화를 관찰하기 위해서 MTT cytotoxicity assay, DAPI staining, TUNEL assay, RT-PCR analysis 방법을 이용하였다. 결과 : 세포독성 검사에서 계혈등 약침액은 자궁경부암 세포주 SNU-17에 농도 의존적으로 세포독성을 나타내었다. 이러한 계혈등 약침액의 세포독성이 세포사별로 인한 것인지 다른 기전에 의한 것인지 알아본 결과 계혈등 약침액에 의한 세포독성은 DAPI staining 과 TUNEL assay에서 세포사멸의 특정적인 소견들을 나타내었다. 계혈등 약침액이 Bax, Caspase-3의 발현에 미치는 영향을 RT-PCR로 관찰한 결과 계혈등 약침액은 Bax, Caspase-3의 발현을 증가시켰다. 결론 : 이상의 결과 계혈등 약침액이 자궁경부암 세포주 SNU-17에서 세포 사멸을 야기하여 자궁경부암의 치료에 유용할 것으로 사료된다.

• Journal of Acupuncture Research
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• v.26 no.3
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• pp.27-38
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• 2009

목적 : 이 연구는 Vipera lebetinat turanica의 사독약침액(蛇毒藥鍼液)(Snake venom toxin, SVT)이 인간 유방암 세포주인 MCF-7과 MDA-MB-231 세포에서 암세포성장의 억제 및 그 기전에 대하여 살펴보고자 하였다. 방법 : SVT를 처리한 후 MCF-7과 MDA-MB-231의 성장억제를 관찰하기 위해 CCK-8 assay를 시행하였고, apoptosis 평가에는 TUNEL assay를 시행하였다. 세포자멸사 관련 세포기전을 보기 위하여 세포내 활성산소량 및 미토콘드리아의 세포막전위 변화를 측정하였고, 세포자멸사 조절 단백인 Bax, Bcl-2 발현 변화 관찰에는 westem blot analysis를 시행하였다. 결과 : MCF-7과 MDA-MB-231 세포에 SVT를 처리한 후, 유방암 세포의 성장, Apoptosis의 유발 및 기전에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. MCF-7 세포와 MDA-MB-231 세포에서 SVT를 처리한 후 유방암 세포 성장이 억제되었다. 2. TUNEL assay를 통한 세포자멸사 평가에서 SVT를 처리한 MCF-7세포와 MDA-MB-231 세포 모두 세포자멸사 활성세포의 유의한 증가를 나타내었다. 3. 세포자멸사 관련 세포기전연구에서 SVT를 처리한 MCF-7 세포와 MDA-MB-231 세포에서 세포내 활성산소의 유의한 증가와 미토콘드리아 세포막 전위의 유의한 변동이 관찰되었다. 4. SVT를 처리한 MCF-7세포와 MDA-MB-231세포는 세포자멸사 관련 단백 발현에서 Bax의 유의한 증가와 Bcl-2의 유의한 감소를 나타내었다. 결론 : 이상의 결과는 SVT가 세포내 활성산소를 증가시킴으로써 미토콘드리아의 세포막전위에 변화를 일으켜 유방암 세포주인 MCF-7과 MDA-MB-231 세포에 세포자멸사를 유발하여 증식억제 효과가 있음을 입증한 것이다.

• The Journal of Internal Korean Medicine
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• v.27 no.2
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• pp.316-326
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• 2006

Backgrounds & Objects: The aim of this study was to investigate the radioprotective effect of Shengmai-san(SMS), a herbal medicine, on mice jejunal crypt cell survival and Apoptosis. Methods: Mice were devided into 4 groups according to radiation dose and SMS treatment: Normal was the group without irradiation. Control was the group treated with D.W before 10 Gy irradiation. SMS 2.9 was sample group treated with 2.9 mg/10 g of SMS extract before 10 Gy irradiation and SMS 29 was sample group treated with 29 mg/10 g of SMS extract before 10 Gy irradiation. And Each group were sacrificedat 24 hours and 72 hours after irradiation. To analyze the crypt survival, hematoxylin-eosin staining was used and to analyze the apoptosis, the TUNEL assay was done. Results: 1. From the microcolony survival assay, the SMS 2.9 and SMS 29 showed the radioprotective effect with a statistical significance compared to the control group at 24 hr (P < 0.01) and 72 hr (p < 0.001) after 10 Gy irradiatien. And the differences of radioprotective effect between SMS 2.9 and SMS 29 were net significant. 2. The results of the TUNEL assay showed that the apoptotic index in SMS 2.9 and SMS 29 was significantly decreased, as compared to the control group at both 24 hr ( p < 0.01) and 72 hr (SMS 2.9 : p < 0.001. SMS 29 : P < 0.01) after 10Gy irradiation And the differences of between SMS 2.9 and SMS 29 were not significant. Conclusions: It could be suggested that the Shengmai-san has a prominent Protective effect in mice intestines against the radiation damage. And the radieprotective effect seems to be related to inhibition of the apoptosis.