Synergistic Effects of Chios Gum Mastic Extract and Low Level Laser Therapy on Osteoblast Differentiation

In the present study, we evaluated the effect of CGM on osteogenic differentiation of cultured osteoblasts, and determined whether combination treatment with LLLT had synergistic effects on osteogenic differentiation. The results indicated that CGM promoted proliferation, differentiation, and mineralization of osteoblasts at the threshold concentration of 10 µg/ml; whereas, CGM showed cytotoxic properties at concentrations above 100 µg/ml. ALP activity and mineralization were increased at concentrations above 10 µg/ml. CGM in concentrations up to 10 µg/ml also increased the expression of osteoblast-activated factors including type I collagen, BMP-2, RUNX2, and Osterix. The CGM (50 µg/ml) and LLLT (80 mW for 15 sec) combination treatment group showed the highest proliferation levels, ALP activity, and mineralization ratios. The combination treatment also increased the levels of phosphorylated forms of p38, ATF2, PKD, ERK, and JNK. In addition, the osteoblast differentiation factors including type I collagen, BMP-2, RUNX2, and Osterix protein levels were clearly increased in the combination treatment group. These results suggested that the combination treatment of CGM and LLLT has synergistic effects on the differentiation and mineralization of osteoblastic cells.

Anticancer Properties of Icariside II in Human Oral Squamous Cell Carcinoma Cells

OSCC is currently the most common malignancy of the head and neck, affecting tens of thousands of patients per year worldwide. Natural flavonoids from plants are potential sources for novel anti-cancer drugs. Icariin is the active ingredient of flavonol glycoside, which is derived from the medical plant Herba Epimedii. A metabolite of icariin, icariside II exhibits a variety of pharmacological actions, including anti-rheumatic, anti-depressant, cardiovascular protective, and immunomodulatory functions. However, the exact mechanism causing the apoptosis-inducing effect of icariside II in OSCC is still not fully understood. In the present study, we assessed the anti-cancer effect of icariside II in OSCC cell lines by measuring its effect on cell viability, cell proliferation, and mitochondria membrane potential (MMP). Icariside II treatment of OSCC cells resulted in a dose- and time-dependent decrease in cell viability. Hoechst staining indicated apoptosis in icariside II-treated HSC cells. Icariside II inhibited cell proliferation and induced apoptosis in HSC cells, with significant increases in all present parameters in HSC-4 cells. The results clearly suggested that icariside II induced apoptosis via activation of intrinsic pathways and caspase cascades in HSC-4 cell lines. The collective findings of the study suggested that Icariside II is a potential treatment for OSCC; in addition, the data could provide a basis for the development of a novel anti-cancer strategy.

Incidence of pterygospinous and pterygoalar bridges in dried skulls of Koreans / 대한해부학회지

Understanding of morphological structures such as the sphenoid spine and pterygoid processes is important during lateral transzygomatic infratemporal fossa approach. In addition, osseous variations such as pterygospinous and pterygoalar bridges are significant in clinical practice because they can produce various neurological disturbances or block the passage of a needle into the trigeminal ganglion through the foramen ovale. Two hundred and eighty-four sides of Korean adult dry skulls were observed to carry out morphometric analysis of the lateral plate of the pterygoid process, to investigate, for the first time among Koreans, the incidence of the pterygospinous and pterygoalar bony bridges, to compare the results with those available for other regional populations, and to discuss their clinical relevance as described on literatures. The mean of maximum widths of the left and right lateral plates of the pterygoid process were 15.99 mm and 16.27 mm, respectively. Also, the mean of maximum heights of the left and right lateral plates were 31.02 mm and 31.01 mm, respectively. The ossified pterygospinous ligament was observed in 51 sides of the skulls (28.0%). Ossification of the pterygospinous ligament was complete in four sides (1.4%). In 47 sides (16.6%), the pterygospinous bridge was incomplete. The ossified pterygoalar ligament was observed in 24 sides of the skulls (8.4%). Ossification was complete in eight sides (2.8%) and incomplete in 16 sides (5.6%). This detailed analysis of the lateral plate of the pterygoid process and related ossification of ligaments can improve the understanding of complex clinical neuralgias associated with this region.

Morphometric Study of the Retromolar Triangle and Foramen in Korean Mandibles / 체질인류학회지

It has been known that the retromolar foramen is a rare anatomic variation observed in the retromolar triangle, a small triangular shaped region posterior to the mandibular third molar. Due to the neurovascular bundle passing through the retromolar foramen, this anatomical structure must be kept in mind during surgical approaches regarding the retromolar area and mandible. Therefore, the authors investigated the morphology of retromolar triangle and the existence and location of retromolar foramen in Korean. And these results were compared with that of other races. We used 308 sides of 154 Korean dry mandibles, unknown gender and age. The retromolar triangle presented predominantly a triangular shape (84.1%), and the maximum height and width were 13.7 mm and 7.1 mm, respectively. In 144 of the 308 sides, the retromolar foramen was observed (46.8%). The existence of the retromolar foramen was seen the same frequency in both sides, and based on a midsagittal line of the retromolar triangle, the retromolar foramen located in more buccal side (75%) than lingual side. The mean distance between the retromolar foramen and the distal edge of the last tooth were found to be 10.3 mm and 6.9 mm, respectively for the second and third molars. According to the present study, the northeast Asians including Korean population show the highest rate of the incidence of the retromolar foramen than other races. The findings suggest that practitioners should take the retromolar foramen into account in surgical procedures involving the retromolar area to protect the patient from the complications such as bleeding or nerve damage.

Apoptotic Effect of Co-treatment with Curcumin and Cisplatin on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line

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 microM of curcumin or 4 microg/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.

Mechanism Underlying Curcumin-induced Apoptosis and Cell Cycle Arrest on SCC25 Human Tongue Squamous Cell Carcinoma Cell Line

Several studies have shown that curcumin, which is derived from the rhizomes of turmeric, possesses antimicrobial, antioxidant and anti-inflammatory properties. The antitumor properties of curcumin have also now been demonstrated more recently in different cancers. This study was undertaken to investigate the modulation of cell cycle-related proteins and the mechanisms underlying apoptosis induction by curcumin in the SCC25 human tongue squamous cell carcinoma cell line. Curcumin treatment of the SCC25 cells resulted in a time- and dose-dependent reduction in cell viability and cell growth, and onset of apoptotic cell death. The curcumin-treated SCC25 cells showed several types of apoptotic manifestations, such as nuclear condensation, DNA fragmentation, reduced MMP and proteasome activity, and a decreased DNA content. In addition, the treated SCC25 cells showed a release of cytochrome c into the cytosol, translocation of AIF and DFF40/CAD into the nuclei, a significant shift in the Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-7, caspase-6, caspase-3, PARP, lamin A/C, and DFF45/ICAD. Furthermore, curcumin exposure resulted in a downregulation of G1 cell cycle-related proteins and upregulation of p27KIP1. Taken together, our findings demonstrate that curcumin strongly inhibits cell proliferation by modulating the expression of G1 cell cycle-related proteins and inducing apoptosis via proteasomal, mitochondrial, and caspase cascades in SCC25 cells.

Apoptotic Effects of Co-Treatment with a Chios Gum Mastic and Eugenol on G361 Human Melanoma Cells

We investigated the synergistic apoptotic effects of co-treatments with Chios gum mastic (CGM) and eugenol on G361 human melanoma cells. An MTT assay was conducted to investigate whether this co-treatment efficiently reduces the viability of G361 cells compared with each single treatment. The induction and augmentation of apoptosis were confirmed by DNA electrophoresis, Hoechst staining, and analyses of DNA hypoploidy. Western blot analysis and immunofluorescent staining were also performed to evaluate expression and translocation of apoptosis-related proteins following CGM and eugenol co-treatment. Proteasome activity and mitochondrial membrane potential (MMP) changes were also assayed.The results indicated that the co-treatment of CGM and eugenol induces multiple pathways and processes associated with an apoptotic response in G361 cells. These include nuclear condensation, DNA fragmentation, a reduction in MMP and proteasome activity, an increase of Bax and decrease of Bcl-2, a decreased DNA content, cytochrome c release 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 40 microg/ml CGM or 300 microM eugenol for 24 hours did not induce apoptosis. Our present data thus suggest that a combination therapy of CGM and eugenol is a potential treatment strategy for human melanoma.

Preferential Cytotoxic Effect of Genistein on G361 Melanoma Cells Via Inhibition of the Expression of Focal Adhesion Kinase

Resistance to the induction of apoptosis is a possible mechanism by which tumor cells can survive anti-neoplastic treatments. Melanoma is notoriously resistant to anti-neoplastic therapy. Previous studies have demonstrated focal adhesion kinase (FAK) overexpression in melanoma cell lines. Given its probable role in mediating resistance to apoptosis, many researchers have sought to determine whether the downregulation of FAK in melanoma cells would confer a greater sensitivity to anti-neoplastic agents. Genistein is a known inhibitor of protein-tyrosine kinase (PTK), which may attenuate the growth of cancer cells by inhibiting the PTK-mediated signaling pathway. This present study was undertaken to investigate the effect of genistein on the expression of FAK and cell cycle related proteins in the G361 melanoma cell line. Genistein was found to have a preferential cytotoxic effect on G361 melanoma cells over HaCaT normal keratinocytes. Genistein decreased the expression of 125 kDa phosphotyrosine kinase and the FAK protein in particular. Genistein treatment did not affect the expression of p53 in G361 cells in which p21 is upregulated. The expression of cyclin B and cdc2 was downregulated by genistein treatment. Taken together, our data indicate that genistein induces the decreased proliferation of G361 melanoma cells via the inhibition of FAK expression and regulation of cell cycle genes. This suggests that the use of genistein may be a viable approach to future melanoma treatments.

HS-1200 Overcomes the Resistance Conferred by Bcl-2 in Human Leukemic U937 Cells

Bcl-2 protects tumor cells from the apoptotic effects of various anti-neoplastic agents. Increased expression of Bcl-2 has been associated with a poor response to chemotherapy in various malignancies, including leukemia. Hence, bypassing the resistance conferred by anti-apoptotic factors such as Bcl-2 represents an attractive therapeutic strategy against cancer cells, including leukemic cells. This study was undertaken to examine whether the anticancer drug, cisplatin and the synthetic chenodeoxycholic acid (CDCA) derivative, HS-1200 show anti-tumor activity in U937 and U937/Bcl-2 cells. Viability assays revealed that HS-1200 overcomes the resistance conferred by Bcl-2 in human leukemic U937 cells. Various apoptosis assessment assays further demonstrated that HS-1200 overcomes the resistance conferred by Bcl-2 in human leukemic U937 cells by inducing apoptosis. In addition HS-1200, but not cisplatin, overcomes the anti-apoptotic effects of Bcl-2 in Bcl-2 over-expressing human leukemic cells (U937/Bcl-2 cells). Notably, we observed that the HS-1200-induced formation of mature promyelocytic leukemia (PML) nuclear bodies (NBs) correlates with a suppression of the anti-apoptotic effects of Bcl-2 in human leukemic cells over-expressing this protein (U937/Bcl-2 cells). Furthermore, HS-1200 was found to induce the association between PML and SUMO-1, Daxx, Sp100, p53 or CBP in the aggregated PML-NBs of U937/Bcl-2 cells. Thus, PML protein and the formation of mature PML-NBs could be considered as therapeutic targets that may help to bypass the resistance to apoptosis conferred by Bcl-2. Elucidating the exact mechanism by which PML regulates Bcl-2 will require further work.

A Case of Branchio-Otic Syndrome / 대한이비인후과학회지

Branchio-oto-renal (BOR) syndrome is a clinically heterogeneous autosomal dominant form of syndromic hearing loss characterized by variable hearing impairment, malformations of the pinnae, the presence of branchial arch remnants, and various renal abnormalities. BOR syndrome is caused by mutations in EYA1 and SIX1, which are critical to organogenesis and are expressed together in developing otic, branchial, and renal tissue. Branchio-otic (BO) syndrome comprises branchial fistulas and preauricular pits, but lacks renal anomalies. We present a case of BO syndrome in 30year-old man with a review of the literature.

Apoptotic Effects of A Cisplatin and Eugenol Co-treatment of G361 Human Melanoma Cells

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 microM eugenol or 3 microM 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.

A Natural Product, Chios Gum Mastic, Induces the Death of HL-60 Cells via Apoptosis and Cell Cycle Arrest

Chios gum mastic (CGM) is produced from Pistiacia lentiscus L var chia, which grows only on Chios Island in Greece. CGM is a kind of resin extracted from the stem and leaves, has been used for many centuries in many Mediterranean countries as a dietary supplement and folk medicine for stomach and duodenal ulcers. CGM is known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis following CGM treatment of HL-60 cells. The viability of the HL-60 cells was assessed using the MTT assay. Hoechst staining and DNA electrophoresis were employed to detect HL-60 cells undergoing apoptosis. Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity analyses were also employed. CGM treatment of HL-60 cells was found to result in a dose- and time-dependent decrease in cell viability and apoptotic cell death. Tested HL-60 cells showed a variety of apoptotic manifestations and induced the downregulation of G1 cell cycle-related proteins. Taken collectively, our present findings demonstrate that CGM strongly induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and also apoptosis via proteasome, mitochondrial and caspase cascades in HL-60 cells. Hence, we provide evidence that a natural product, CGM could be considered as a novel therapeutic for human leukemia.

Mechanism Underlying NaF-induced Apoptosis and Cell Cycle Arrest on G361 Human Melanoma Cell Line / 체질인류학회지

Fluoride is widely used in dentistry to prevent dental caries, even though the safety of fluoride is a controversial issue. There are no known adverse effects of long-term fluoride ingestion for caries prevention, but an overdose can cause serious acute toxicity. Nevertheless it is accepted that fluoride is an important material for oral health. This study was undertaken to investigate the modulation of cell cycle-related proteins and apoptosis induction underlying mechanism by NaF treatment on G361 human melanoma cell line. The viability of G361 cells and the growth inhibition of G361 cells were assessed by MTT assay and clonogenic assay respectively. Hoechst staining, DNA electrophoresis and TUNEL staining were conducted to observe G361 cells undergoing apoptosis. G361 cells were treated with NaF, and Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, MMP activity and proteasome activity were performed. NaF treatment in G361 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. And tested G361 cells showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA contents, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, a significant shift of Bax/Bcl-2 ratio, and the activation of caspase-9, caspase-7, caspase-6, caspase-3, PARP, Lamin A/C and DFF45 (ICAD). Furthermore, NaF resulted in down-regulation of the G1 cell cycle-related proteins, and up-regulation of p53. Taken collectively, our present findings demonstrate that NaF strongly inhibits cell proliferation by modulating the expression of the G1 cell cycle-related proteins and induces apoptosis via proteasome, mitochondria and caspase cascades in G361 cells.

Clinical and anatomical approach using Sihler's staining technique (whole mount nerve stain) / 대한해부학회지

Sihler's staining allows visualization of the nerve distribution within soft tissues without extensive dissection and does not require slide preparation, unlike traditional approaches. This technique can be applied to the mucosa, muscle, and organs that contain myelinated nerve fibers. In particular, Sihler's technique may be considered the best tool for observing nerve distribution within skeletal muscles. The intramuscular distribution pattern of nerves is difficult to observe through manual manipulation due to the gradual tapering of nerves toward the terminal end of muscles, so it should be accompanied by histological studies to establish the finer branches therein. This method provides useful information not only for anatomists but also for physiologists and clinicians. Advanced knowledge of the nerve distribution patterns will be useful for developing guidelines for clinicians who perform operations such as muscle resection, tendon transplantation, and botulinum toxin injection. Furthermore, it is a useful technique to develop neurosurgical techniques and perform electrophysiological experiments. In this review, Sihler's staining technique is described in detail, covering its history, staining protocol, advantages, disadvantages, and possible applications. The application of this technique for determining the arterial distribution pattern is also described additionally in this study.

Distribution of Histologic Type of Nasal Polyp and Expression of Vascular Endothelial Growth Factor According to Nasal Polyp Type / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: To determine the frequency of the histologic types of nasal polyp in Korea and their relationships with respect to age, laterality, asthma, allergic rhinitis (AR) and expression of vascular endothelial growth factor (VEGF). SUBJECTS AND METHOD: Tissue slides obtained from 282 patients with nasal polyps were examined; polyps were classified either as eosinophilic polyp or chronic inflammatory polyp. VEGF expression was determined using immunohistochemical staining. RESULTS: Of the 282 subjects, 169 (59.9%) had chronic inflammatory polyps, 113 (40.1%) had eosinophilic polyps, and 232 (82.3%) had bilateral polyps. Twenty-two subjects (7.8%) had asthma and 23 (8.2%) had AR. There was no statistical relationship between nasal polyp type and laterality or the presence of asthma or AR. Of 10 children, 9 (90%) had chronic inflammatory polyps. VEGF expression was significantly higher in eosinophilic polyps than in chronic inflammatory polyps, and significantly higher in the samples of each polyp type from the subjects with AR than those without AR. In subjects with asthma, however, the VEGF expression did not differ between eosinophilic polyps and chronic inflammatory polyp samples. CONCLUSION: In the Korean population, chronic inflammatory nasal polyps are more common than eosinophilic nasal polyps. VEGF expression was the highest in eosinophilic polyps of the subjects with AR, suggesting that VEGF might contribute to the polyp formation via local allergic action.

Anatomy of the Blood Supply of Dorsal Side of Thumb and First Web / 체질인류학회지

Distal thumb injuries are a common and difficult problem for hand surgeons. Coverage of soft tissue on the fingers may be difficult due to the size of the defect or the limitation of local flap mobilization. However, the variable anatomy of the dorsal hand vascular system sometimes prevents successful flap harvest. The purpose of this study was to clarify the vascular anatomy of the dorsal side of the thumb and the first web for the flaps. Twenty six hands (13 right and 13 left hands) from Korean embalmed cadavers were dissected. A catheter was inserted into the radial artery in the forearm, and the red colored latex (Latex 671, Dupont Industry, France) was injected until the dorsum of the hand was colored. The arrangement of the first dorsal metacarpal artery (FDMA) and its branches were vary and classified into three categories according to their branching patterns; Both dorso-ulnar thumb branch (DUTB) and dorso-radial index branch (DRIB) arose from the FDMA (10 cases, 38.5%). Each DUTB and DRIB arose separately from the radial artery (5 cases, 19.2%). The DUTB and the DRIB originated from the princeps pollicis artery and the radial artery, respectively (11 cases, 42.3%). The typical course of the FDMA and its branches ran overlying the first dorsal interosseous muscle in 17 cases of the 26 specimens (65.4%). However, in nine cases (34.6%) the DRIB ran on the first dorsal interosseous muscle and the DUTB had a deep course within the substance of the first dorsal interosseous muscle. The FDMA flap represents a good option to cover defects for the thumb. These anatomical findings in the present study could provide useful knowledge of flaps for dorsal aspect of the thumb and the first web.

Apoptotic Effect of Co-treatment with Chios Gum Mastic and HS-1200 on G361 Human Melanoma Cell Line / 대한해부학회지

Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. Recently it reported that CGM induce apoptosis in a few cancer cells in vitro. Bile acids and their synthetic derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. It has been reported that the synthetic chenodeoxycholic acid (CDCA) derivatives showed apoptosis-inducing activity on various cancer cells in vitro. This study was undertaken to investigate the synergistic apoptotic effect of cotreatment with a natural product, CGM and a CDCA derivative, HS-1200 on G361 human melanoma cells. To investigate whether the co-treatment of CGM and HS-1200 compared with each single treatment efficiently reduced the viability of G361 cells, MTT assay was conducted. To investigate augmentation of apoptosis in G631 cells co-treated with CGM and HS-1200, DNA electrophoresis, Hoechst staining, proteasome activity assay, flow cytometry, Westen blot analyses, immunofluorescent staining and confocal microscopy were performed. In this study, G361 cells co-treated with CGM and HS-1200 showed several lines of apoptotic manifestation such as nuclear condensations, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, activation of caspase-9, caspase-3, PARP and DFF45 (ICAD), and up-regulation of Bax whereas each single treated G361 cells did not. Although the single treatment of 40 micro/mL CGM or 25 micro HS-1200 for 24 hrs did not induce apoptosis, the co-treatment of them induced prominently apoptosis. Therefore, combination therapy of CGM and HS-1200 could be considered, in the future, as an alternative therapeutic strategy for human melanoma.

Chios Gum Mastic Induces Cell Cycle Arrest and Apoptosis in Human Osteosarcoma Cells / 대한해부학회지

Chios gum mastic (CGM) is a resin produced from the stem and leaves of Pistiacia lentiscus L var chia, a plant which grows only on Chios Island in Greece. CGM has been used for many centuries as a dietary supplement and folk medicine for stomach and duodenal ulcers in many Mediterranean countries and is also known to induce cell cycle arrest and apoptosis in some cancer cells. This study was undertaken to investigate the alteration of the cell cycle and induction of apoptosis by CGM treatment on human osteosarcoma (HOS) cells. The viability and the growth inhibition of HOS cells were assessed by the MTT assay and clonogenic assay respectively. The hoechst staining, TUNEL assay and DNA electrophoresis were conducted to observe the HOS cells undergoing apoptosis. HOS cells were treated with CGM, and Western blotting, immunocytochemistry, confocal microscopy, FACScan flow cytometry, mitochondrial membrane potential change and proteasome activity were conducted. CGM treatment of HOS cells was found to result in a dose- and time-dependent decrease in cell viability, a dose-dependent inhibition of cell growth, and apoptotic cell death. Tested HOS cells also showed several lines of apoptotic manifestation and G1 arrest in cell cycle progression. In summary, this study clearly demonstrated that CGM induces G1 cell cycle arrest via the modulation of cell cycle-related proteins, and apoptosis via proteasome, mitochondrial and caspase cascades in HOS cells. Therefore, our data provide the possibility that a natural product, CGM could be considered as a novel therapeutic strategy for human osteosarcoma.

A Synthetic Chenodeoxycholic Acid Derivative, HS-1200-induced Apoptosis of RBL-2H3 Cells / 대한해부학회지

Bile acids and synthetic bile acid derivatives induced apoptosis in various kinds of cancer cells and anticancer effects. Although synthetic chenodeoxycholic acid (CDCA) derivatives have been demonstrated to induce apoptosis of various cancer cells, there is no report on their effect on RBL-2H3 basophilic leukemia cell line to date. Therefore, this study was undertaken to examine in vitro effects of cytotoxicity and growth inhibition, and the molecular mechanism underlying induction of apoptosis in RBL-2H3 cells treated with a synthetic CDCA derivative, HS-1200. The viability and the growth inhibition of RBL-2H3 cells were assessed by MTT assay and clonogenic assay respectively. The Hoechst staining and DNA electrophoresis were conducted to observe RBL-2H3 cells undergoing apoptosis. RBL-2H3 cells were treated with HS-1200, and Western blotting, immunocytochemistry, confocal microscopy, DNA hypoploidy assay, MMP activity and proteasome activity were performed. HS-1200 treatment of RBL-2H3 cells resulted in a time- and does-dependent decrease of cell viability and a does-dependent inhibition of cell growth, and induced apoptotic cell death. Furthermore, HS-1200 treatment result in the alteration of G1 cell cycle-related proteins. And tested RBL-2H3 cells showed several lines of apoptotic manifestation.We presented data indicating that HS-1200 induces apoptois via the proteasome, mitochondria and caspase pathway, and induces the alteration of the G1 cell cycle-related proteins in RBL-2H3 cells. Therefore our data provide the possibility that HS-1200 could be as a novel therapeutic strategy in the allergy treatment.

Apoptotic Effect of Co-Treatment with a Natural Product, Chios Gum Mastic, and a Proteasome Inhibitor, Lactacystin, on Human Osteosarcoma Cells / 대한해부학회지

Chios gum mastic (CGM) is a resinous exudate obtained from the stem and the main leaves of Pistacia lenticulus tree native to Mediterranean areas. Recently, it was reported that CGM induced apoptosis in a few cancer cells in vitro. Since recent studies indicated the synergistic interactions between the apoptotic stimulus and a proteasome inhibitor, the ubiquintin-proteasome pathway has become an attractive target in cancer therapy. And to date, there has been no report of the synergistic apoptotic effect between CGM and a proteasome inhibitor to become an attractive target in cancer therapy. Therefore, this study was undertaken to investigate the synergistic apoptotic effect of co-treatment with a natural product, CGM, and a proteasome inhibitor, lactacystin, on human osteosarcoma (HOS) cells. To investigate whether the co-treatment of CGM and lactacystin compared with each single treatment efficiently induced apoptosis on HOS cells, MTT assay, DNA electrophoresis, Hoechst staining, DNA hypoploidy assay, Westen blot analysis, immunofluorescent staining, proteasome activity and mitochondrial membrane potential (MMP) change were performed. In this study, HOS cells co-treated with CGM and lactacystin showed several lines of apoptotic manifestation such as nuclear condensation, DNA fragmentation, the reduction of MMP and proteasome activity, the decrease of DNA content, the release of cytochrome c into cytosol, the translocation of AIF and DFF40 (CAD) onto nuclei, and activation of caspase-7, caspase-3, PARP and DFF45 (ICAD) whereas each single treated HOS cells hardly showed. We presented data indicating that the co-treatment of CGM and lactacystin induced potentially apoptosis whereas each single treatment did slightly. Moreover, the co-treatment of CGM and lactacystin potentiated the inhibition of proteasome activity. Therefore, our data provide the possibility that combination therapy of CGM and lactacystin could be considered as a novel therapeutic strategy for human osteosarcoma.