• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.186-198
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• 2006

Purpose : The purpose of this study is to demonstrate the direct inhibitory effect of Hyulbuchukeotang on the proliferation of uterine leiomyoma cells through an experiment treating uterine leiomyoma cells cultivated by explantation with indicated concentrations of Hyulbuchukeotang and to research the gene expression related to cell cycle ill order to discover the connection with apoptosis and its mechanism by analyzing cell cycle. Methods : After primary culture of uterine leiomyoma cells, the cultivated uterine leiomyoma cells were treated with indicated concentrations of Hyulbuchukeotang for 24 hours. The inhibitory effect on the cell proliferation was determined by the cell count assay. The value of a cell count assay represent the percentage of cells in a phase of the cell cycle compared with total cells. In addition, a link between Hyulbuchukeotang and apoptosis was examined through flow cytometric analysis by FACS and DNA fragmentation analysis. Finally, the degree of gene expression related to cell cycle was evaluated by Western blot analysis. Results : The inhibitory effect of Hyulbuchukeotang increase of uterine leiomyoma cells treated with indicated concentrations of Hyulbuchkeotang increases. The result of gene expression related to G1 phase after treating with 100, 250, 500, 1,000 ${\mu}g/ml$ concentrations of Hyulbuchukeotang. on uterine leiomyoma cells is that the gene expression of p27 was increased but that of p53 an p21 remained unchanged and the gene of pRB, pro-caspase 3 was decreased. Conclusion Through the mentioned experiments, it is demonstrated that Hyulbuchkeotang is effective in inhibiting Proliferation of uterine leiomyoma cells by extending cell cycle G1. However it is not considered that the inhibitory effect results from the aptoposis.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.2
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• pp.25-42
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• 2007

Purpose : Uterine leiomyoma (fibroids) are benign smooth muscle tumors originating from the myometrium. These benign neoplasms of monoclonal origin are typically diagnosed during the reproductive years, occurring only after puberty and tending to regress after menopause. In the present study we used Chiljehyangbu-hwan to determine its growth inhibitory effect and apoptosis in human uterine leiomyoma cells. Methods : Primary cultured human uterine leiomyoma cells were treated with Chiljehyangbu-hwan. Cell viability analysis was analyzed by MTS assay and FACS was performed to ascertain the effects Chiljehyangbu-hwan. DNA fragmentation analysis and casapase-3 activity test were done. Expression of apoptosis related proteins were evaluated by Western blot analysis. Results : Cell viability was significantly influenced by Chiljehyangbu-hwan treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. FACS showed that Chiljehyangbu-hwan induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increased expression in Bid and Bax were observed. Chiljehyangbu-hwan treatment of uterine leiomyoma cells resulted in a concentration-dependent cell death induced via the mitochondrial pathway.

• The Journal of Korean Obstetrics and Gynecology
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• v.20 no.3
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• pp.1-12
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• 2007

Purpose: Uterine leiomyomas (fibroids) are common estrogen-dependent uterine tumors. Houttuynia cordata thunberg has cancer-preventing properties and often used in Chinese medicine. In the present study we used Houttuynia cordata thunberg to determine its effect on cell proliferation and apoptosis in human uterine leiomyoma cells. Methods: Primary cultured human uterine leiomyoma cells were treated with Houttuynia cordata thunberg. Cell viability analysis was analyzed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTS) assay and Flow cytometry was performed to ascertain the effects Houttuynia cordata thunberg. Expression of cell cycle related proteins and apoptosis related proteins were evaluated by Western blot analysis. Results: Cell viability was significantly influenced by Houttuynia cordata thunberg treatment in a dose-dependent manner in leiomyoma cells compare to normal myometrial cells. Flow cytometric analysis showed that Houttuynia cordata thunberg induced Sub G1 arrest. DNA fragmentation assay was carried out and apoptosis was detected. Activation of caspase-3, down-regulation of Bcl-2, with concomitant increase in p21 was observed. Houttuynia cordata thunberg treatment of uterine leiomyoma cellsresulted in a concentration-dependent cell death induced via the caspase dependent mechanism. Conclusion: These results suggest that Houttuynia cordata thunberg treatment in uterine leiomyoma cells leads to growth inhibition and induced apoptosis. These results suggest that Houttuynia cordata thunberg will be a promising agent for use in therapeutics agents against human uterine endometrial cancer.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.2
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• pp.199-213
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• 2006

Purpose : This study was aimed to investigate the inhibitory effect of Sparganii Rhizoma on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : This study was evaluated the number of death cells treated with indicated concentration of Sparganii Rhizoma and investigated cell death rate by MTS assay. Furthermore, fluorescence-activated cell sorter analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results :1) The inhibitory effect on the growth of uterine leiomyoma cell treated with Sparganii Rhizoma was increased in a dose dependent manner. 2) As the result of FACS analysis, subG1 phase incrase was observed 23.49% inuterine leiomyoma cell treated with Sparganii Rhizoma at $500\;{\mu}g/ml$ compared to control.. 3) The gene expression of p53, p21 related cell apoptosis was increased according to increasing concentration but p27 was none exchanged. 4) The expression of cyclin A, D and E was decreased in a concentration proportional and then the dephosphorylation of pRb was increased. 5) The character of apoptosis, DNA fragmentation was significantly observed according to increasing concentration. 6) The expression of pro-caspase3 were decreased dependent on treatment concentration and activated PARP took place. Conclusion : The inhibitory effect of Sparganii Rhizoma on the proliferation of human uterine leiomyoma cells was observed with apoptosis and cell cycle arrest. These data suggest that Sparganii Rhizoma might be candidate of medical therapy for uterine leiomyoma.

• Journal of Physiology & Pathology in Korean Medicine
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• v.21 no.1
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• pp.158-164
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• 2007

Uterine leiomyoma is the most common tumor in the female genital tract. Although the tumor is benign, it is a matter of paramount importance since it often causes profuse menstrual bleeding, pressure symptoms and infertility. Nevertheless, the etiology and pathophysiology of this abnormality remain poorly understood. The traditional definitive treatment for uterine leiomyomas is hysterectomy and, even today, symptomatic leiomyomas are the leading cause of hysterectomy in Korea. Clearly, the development of a safe, effective, and nonsurgical method of treatment for leiomyoma would be of great benefit to many women. This study demonstrated growth inhibition of uterine leiomyoma cells using Haeohyultang (HT). When human leiomyoma cells were treated with Haeohyultang, cells showed dose-dependent growth inhibitory effect. Cell growth was inhibited by over 40% as determined by both cell counts and MTS assay. Reduction of cellular viability as a consequence of exposure to Haeohyultang resulted from induction of apoptosis, as assessed by DNA fragmentation, PARP cleavage, caspase 9 and caspase 3 assay. Flow cytometry analysis with uterine leiomyoma cells demonstrated sub G1 cell cycle arrest after treatment with drug Haeohyultang. But, the expression levels of p27 and p21 were not changed in Haeohyultang treated cells compared with control. However, the expression levels of clAP1 were reduced by Haeohyultang compared with control. This reduction of clAP1 data means activation of the caspase family, and then induction of PARP cleavage and apoptosis. These results suggest that Haeohyultang may be potential therapeutic approach in the clinical management of uterine leiomyoma.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.1-12
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• 2006

Purpose : This study was aimed to investigate the inhibitory effect of Leonurus sibiricus on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated concentration of Leonurus sibiricus and investigated cell death rate by MTS assay. Furthermore, flow cytometry analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis and then we observed the differential gene expression by western blot analysis. Results : Leonurus sibiricus significantly inhibited the proliferation of uterine leiomyoma cell in a dose-dependent and time dependent manner. Fluorescence activated cell sorter (FACS) analysis indicated that Leonurus sibiricus induced G1 cell cycle arrest. Leonurus sibiricus enhanced the expression of p27 and p53 with cell cycle arrest. Conclusion : These findings suggest that Leonurus sibiricus is a candidate agent for the treatment of uterine leiomyoma. p27, $p53^{1}$ may play an important role in Leonurus sibiricus-induced cell cycle arrest and cell growth inhibition.

• The Journal of Korean Obstetrics and Gynecology
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• v.18 no.1
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• pp.181-191
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• 2005

Purpose : ${\beta}$-sitosterol is kind of phytosterols or plant which are structurally similar to cholesterol. This study was aimed to investigate the inhibitory effect of the ${\beta}$-sitosterol on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated time of the ${\beta}$-sitosterol and investigated cell death rate by cell count assay. Furthermore, flow cytometry analysis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the growth of uterine leiomyoma cell treated with the ${\beta}$-sitosterol $16{\mu}M$ was increased in a time dependent. 2) The result of flow cytometry analysis, subG1 phase arrest related cell apoptosis was investigated 16.97% in uterine leiomyoma cell treated with the ${\beta}$-sitosterol $16{\mu}M$ and showed the fashion of proportional time dependent. 3) The gene expression of p27, p21 related cell cycle was increased according to increasing time interval but cyclin E-CDK2 complex was decreased expression. 4) The character of apoptosis, DNA fragmentation was significantly observed on the time dependent. 5) The expression of pro-caspase 3 and PARP were decreased dependent on treatment with time dependent. Conclusion : This study showed that the ${\beta}$-sitosterol have the inhibitory effect on the proliferation of human uterine leiomyoma cell and the effect was related with apoptosis.

• The Journal of Korean Obstetrics and Gynecology
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• v.19 no.3
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• pp.135-149
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• 2006

Purpose : This study was aimed to investigate the inhibitory effect of Millettia Reticulatas on the proliferation of human uterine leiomyoma cells and the expression of gene related the mechanism of cell apoptosis. Methods : We counted the number of death cells treated with indicated concentration of Millettia Reticulatas and investigated cell death rate by MTS assay. Furthermore, flow cytometry analyis and DNA fragmentation assay were used to dissect between necrosis and apoptosis. and then we observed the differential gene expression by western blot analysis. Results : 1) The inhibitory effect on the growth of uterine leiomyoma cell treated with Millettia Reticulatas was increased in a concentration proportional. 2) The result of flow cytometry analysis. subG1 phase arrest related3 cell apoptosis was investigated 23.49% in uterine leiomyoma cell treated Millettia Reticulatas and showed the fession of proportional concentration. 3) The gene expression of p27, p53, p21, p16 related cell cycle was increased according to increasing concentration but cyclin E was none exchanged. 4) The character of apoptosis, DNA fragmentation was significantly observed the fession of proportional concentration. 5) The expression of pro-caspase3 and PARP were decreased dependent on treatment concentration. Conclusion : This study showed that Millettia Reticulatas have the inhibitory effect on the proliferation of human uterine leiomyoma cell and the effect was related with apoptosis. The apoptotic mechanism was observed that the gene expression of p27, p53, p21, p16 related cell cycle was increased according to increasing treatment concentration, induced G1 phase arrest and finally cell death was occurred. The decreased expression of pro-caspase 3 and PARP were noted that apoptosis was related with caspase pathway.

• Korean Journal of Veterinary Pathology
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• v.5 no.2
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• pp.71-74
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• 2001

Masses of the uterine cervix were detected from two dogs, being a 2-year-old female Yorkshire tarrier and a 12-year-old female Basset Hound. Grossly, the masses measuring 3.O$\times$3.O$\times$4.0 cm and 12.O$\times$l2.O$\times$10.07m were prominent, sharply circumscribed, fleshy tumors in the uterine cervix. Histologically, the masses consisted of smooth muscle cells interwoven in bundles, some of which were cut longitudinally (elongated nuclei) and others transversely. Tumor cell nuclei were ordinarily cigar shaped and had rounded blunt ends in the longitudinal plane. There were low mitotic figures without abnormal ones. From these results, these cases were diagnosed as leiomyoma of the uterine cervix. To our Knowledge, no similar tumors have been reported in the uterine cervix.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.1
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• pp.200-205
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• 2004

Uterine leiomyoma is the most common tumor in the female genital tract. Although the tumor is benign, it is of paramount importance since it often causes profuse menstrual bleeding, pressure symptoms, and infertility. Nevertheless, the etiology and patholphysiology of this abnormality remain poorly understood. The traditional definitive treatment for uterine leiomyomas is hysterectomy and, even today, symptomatic leiomyomas are the leading cause of hysterectomy in Korea. Clearly, the development of a safe, effective, and nonsurgical method of treatment for leiomyoma would be of great benefit to many women. The present study was designed to investigate the effect of Rhubarb on apoptosis in uterine leiomyoma cells. Results demonstrate that Rhubarb inhibited cell growth in dose-dependent manner. Cell growth significantly decreased to 60% of control in the treatment of Rhubarb (300㎍/㎖). Associated with the decreased response, there was a concomitant and significant delay of subG1 8.32% above baseline in the treatment of Rhubarb (300㎍/㎖). The delay of subG1 showed a dose-dependent manner, as evidenced by the flow cytometry. The reduced cellular viability on exposure to Rhubarb may represent the induction of apoptosis, at least in part, as concomitantly evidenced by enhanced DNA fragmentation, PARP cleavage and caspase 9 and decreased pro-caspase 3. In addition, Rhubarb decreased clAP1 expression levels in dose-dependent manner. Talcen together, there results suggest that Rhubarb can produce a potent inhibition effect of apoptosis and implicate the delay of G1 phase in the cell cycle and pathways of caspase 3 and 9 in the mechanism underlying inhibitory apoptosis effect of Rhubarb.