• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.3
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• pp.819-825
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• 2003

pharbitis nil and Taraxacum mongolicum are representative herbs that have been used for cancer treatment in Korean traditional medicine. To understand the molecular basis of the antitumor function, we analyzed the effect of these herbs on proliferation and apoptosis of tumor cells using a gastric cancer cell line AGS. Cell counting assay showed that pharbitis nil strongly inhibit cell proliferation Of AGS whereas Taraxacum mongolicum exhibit no detectable effect on cellular growth. [³H]thymidine uptake analysis also demonstrated that DNA replication of AGS is suppressed in a dose-dependent manner by treatment with pharbitis nil. Additionally, tryphan blue exclusion assay showed that Pharbitis nil induce apoptotic cell death of AGS in a dose-dependent. To explore whether anti antiproliferative and/or proapototic property of Pharbitis nil is associated with their effect on gene expression, we performed RT-PCR analysis of cell cycle- and apoptosis-related genes. Interestingly, mRNA expression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly reduced by Pharbitis nil. Taraxacum mongolicum also showed inhibitory action on expression of these growth-promoting protooncogene but there effects are less significant, as compared to Pharbitis nil. Furthermore, it was also found that Pharbitis nil activates expression of the p53 tumor suppressor and its downstream effector p21Waf1, which induce G1 cell cycle arrest and apoptosis. Collectively, our data demonstrate that Pharbitis nil induce growth inhibition and apoptosis of human gastric cancer cells and these effects are accompanied with down-and up-regulation of growth-regulating protooncogenes and tumor suppressor genes, respectively. This observation thus suggests that the anticancer effect of Pharbitis nil might be associated with its regulatory capability of tumor-related gene expression.

• The Journal of Internal Korean Medicine
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• v.24 no.1
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• pp.134-143
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• 2003

Objectives : We performed this study to understand the molecular basis of the antitumor effect of Saussurea lappa, Pharbitis nil, Plantago asiatica and Taraxacum mongolicum, which have been used for cancer treatment in Korean traditional medicine. Design: We analyzed, the effect of these medicinal herbs on proliferation and apoptosis of tumor cells and its association with gene expression, We performed semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) analysis of cell cycle- and apoptosis-related genes using a gastric cancer cell line AGS. Results : Cell counting assay and $[^3H]thymidine$ uptake analysis showed that Saussurea lappa and Pharbitis nil strongly inhibit cell proliferation of AGS in a dose-dependent manner. Interestingly, gene espression assay revealed that mRNA espression levels of c-Jun, c-Fos, c-Myc, and Cyclin D1 were markedly decreased by Saussurea lappa and Pharbitis nil. Furthermore, Saussurea lappa was identified to activate expression of the p53 tumor suppressor and its downstream effector $p21^{Wafl}$, which leads to $G_1$ cell cycle arrest and apoptosis. These observations suggest that the anticancer effect of Saussurea lappa and Pharbitis nil might be associated with their regulatory capability of tumor-related gene expression.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.6
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• pp.1843-1849
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• 2004

Conventional medicines have usually sorted to a number of treatments such asoperation, radiotherapy, and chemotherapy. The existing anti-cancer agents, designed to eradicate cancer cells, have strong toxicities, also with leading to harmful side effects. Recently, a number of researches on natural products have been actively carried out in efforts to develop new treatments that can decrease side effects or increase anti-cancer effects. We performed this study to understand the molecular basis underlying the antitumor effects of Pharbitis nil, and Plantago asiatica, which have been used for herbal medicinal treatments against cancers in East Asia. We analyzed the effects of these medicinal herbs on proliferation and on expression of cell growth/apoptosis related molecules, with using an AGS gastric cancer cell line. The treatment of Pharbitis nil dramatically reduced cell viabilities in a dose and time-dependent manner, but Plantago asiatica didn't. FACS analysis and Annexin V staining assay also showed that Pharbitis nil induce apoptotic cell death of AGS. Expression analyses via RT-PCR and Western blots revealed that Pharbitis nil didn't increase expression of the p53 and its downstream effector p21/sup wafl/, and that the both increased expression of apoptosis related Sax and cleavage of active caspase-3 protein. We also confirmed the translocation of Sax to mitochondria. Collectively, our data demonstrate that Pharbitis nilinduce growth inhibition and apoptosis of human gastric cancer cells, and these effects are correlated with down- and up-regulation of growth-regulating apoptotic and tumor suppressor genes, respectively.

• Journal of Plant Biology
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• v.25 no.4
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• pp.169-174
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• 1982

Using double immunodiffusion and immunoelectrophoretic techniques, attempts were made to detect any protein changes in leaf tissues of a short-day plant, Pharbitis nil Chois. variety Violet during floral induction under 8 hr light, 16 hr dark cycles. Immunoprecipitin systems shwoed at least four proteins newly appeared in the induced leaf tissues. Accumulation of the proteins were observed as the induction proceeded.

• Journal of Photoscience
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• v.12 no.1
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• pp.1-9
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• 2005

The ubiquitin E3 ligase COP1 (Constitutive Photomorphogenesis 1) is a protein repressor of photomorphogenesis in Arabidopsisplants, and it found in various organisms, including animals. The COP1 protein regulates the stability of many of the light-signaling components that are involved in photomorphogenesis and in the developmental processes. To study the effect of COP1 on flowering in a short day plant, we have cloned a full-length of PnCOP1 (Pharbitis nil COP1) cDNA from Pharbitis nil Choisy cv. Violet, and we examined its transcript levels under various conditions. A full-length PnCOP1 cDNA consists of 2,280 bp nucleotidesthat contain 47 bp of 5'-UTR, 232 bp of 3'-UTR including the poly (A) tail, and 1,998 bp of the coding sequence. The deduced amino acid sequence contains 666 amino acids, giving it a theoretical molecular weight of 75 kD and a isolectric point of 6.2. The PnCOP1 contains three distinct domains, an N-terminal $Zn^2+$-binding RING-finger domain, a coiled-coil structure, and WD40 repeats at the C-terminal, implying that the protein plays a role in protein-protein interactions. The PnCOP1 transcript was detected in the cotyledon, hypocotyls and leaves, but not in root. The levels of the PnCOP1 transcript were reduced in leaves that were a farther distance away from the cotyledons. The expression level of the PnCOP1 gene was inhibited by light, while the expression was increased in the dark. During the floral inductive 16 hour-dark period for Pharbitis nil, the expression was increased and it reached its maximum at the 12th hour of the dark period. The levels of PnCOP1 mRNA were dramatically reduced upon light illumination. These results suggest that PnCOP1 may play an important function in the floral induction of Pharbitis nil.

• Proceedings of the PSK Conference
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• 2002.10a
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• pp.368.3-369
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• 2002

Pharbitis nil Choisy (convolvulaceae) is an annual vine plant and grows at the wayside of Korea. Japan. China and India. The seeds of blue or red Pharbitis nil Choisy. Pharbitidis Semen, is black or red-brown. This seeds have been used as a purgative. From a preliminary experiment. Pharbitidis Semen exhibited anti-cancer activity. MeOH extract of this seeds was subsequently fractionated into four parts: methylene chloride, ethylacetate, n-butanol and water fractions. (omitted)

• Journal of Plant Biology
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• v.38 no.3
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• pp.227-234
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• 1995

Exogenous putrescine of 0.5 mM or higher concentratons applied during a 16 h inductive dark period could elevate putrescine content in cotyledons of Pharbitis nil Choisy cv. Violet, a short-day plant, resulting in complete blocking of photoperiodic floral induction. Titers of putrescine, spermidine and spermine in the cotyledons were traced throughout a 16 h dark period. While non-induced cotyledons under continous light slightly increased levels of polyamines, induced tissue maintaiend its putrescine, spermidine and spermine levels as low as 66.4%, 60.9% and 84.9% of non-induced levels respecitvely. Endogenous polyamines kept at lower levels in the inductive dark period were found to upsurge by a night break treatment of 10 min light in the middle of the dark and consequently the inductive dark effect was canceled. Elevation of polyamine titers could also be induced by 100 $\mu$L/L ethylene treatment which completely suppressed floral induction. Compared to untreated cotyledons, ehtylene-treated tissues increased putrescine content by as much as 136.5% in 12 h and spermidine level by up to 130.1% in 8 h. Ethylene-treated cotyledons not only increased endogenous polyamine content but also liberate ethylene in the second half of the inductive dark period accumulating up to three to fourfold level supporting a hypothesis that ethylene-treated tissues are stimulated to produce ethylene which in turn accelerates polyamine biosynthesis in the tissues. It is postulated that substantially low polyamine titers in the inductive dark period would be one of the necessary factors controlling photoperiodic induction of flowering in Pharbitis nil and the inhibitory effects of night break and exogenous ethylene treatment may be atributed to their action to stimulate endogenous polyamine production.

• Journal of Korean Medicine Rehabilitation
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• v.21 no.1
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• pp.47-56
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• 2011

Objectives : This study was conducted to identify the antimicrobial anti-oxidative and tyrosinase inhibitory effect of three kinds of herbal plants, Polygonum cuspidatum, Chelidonii herba, Pharbitis nil L. naturally grown across the nation. Methods : To investigate in vitro anti-oxidative activity assay, antimicrobial effect and tyrosinase inhibitory activity, MeOH 80% extract, n-hexane, chloroform, ethyl acetate, n-butanol and water fractions of herbal plants were tested by diphenylpicrylhydrazyl(DPPH) method. Results : Bsutanol fractions of each plants showed about 90% on anti-oxidative effect. In case of tyrosinase activity, ethlyacetate fraction of Polygonum cuspidatum showed a potent inhibition effect. In the antimicrobial effect, ethylacetate fraction of Polygonum cuspidatu, hexane fractions of Chelidonii herba and the methanol 80% extracts of Pharbitis nil L. exhibited inhibition effect significantly. Conclusions : In MeOH 80% extracts, we couldn't found any extracts or fractions which have antimicrobial, anti-oxidative and tyrosinase inhibitory activity, however ethylacetate fractions of Polygonum cuspidatum showed effects commonly in these three assay system.

• Journal of Plant Biology
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• v.39 no.4
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• pp.257-263
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• 1996

Flower-inducing activity in the phloem exudata of Pharbitis cotyledons was investigated using the bioassay of Pharbitis and Lemna. By SDS-PAGE and 2-D gel electrophoresis of the phloem exudate, two polypeptides of 11 kDa and of approximately 32 kDa (pI 6.9) showing qualitative changes during the flower induction were detected. A polypeptide of approximately 20 kDa (pI 4.8) specifically labeled in vivo with [35S]methionine was found during the inductive dark period in Pharbitis cotyledon tissues. The polypeptide of the equivalent molecular mass and with the identicl pI value was also detected by in vitro translation assay. Thus, it is assumed that the 20 kDa polypeptide plays a role in the process of flower induction in Pharbitis cotyledons.

• Journal of Photoscience
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• v.5 no.3
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• pp.131-135
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• 1998

Using differential display reverse transcription technique, the present study attempted to isolate and characterize genes specifically expressed in cotyledons of Pharbitis nil Choisy cv. Violet during floral induction. A total of 107 bands specific to the inductive condition were initially obtained with 80 primer sets of 20 different arbitrary primers combined with 4 kinds of T12MN. In northern blot analysis with reamplified cDNAs as probes, three cDNAs were detected to be expressed specificcally in the induced cotyledon tissues, and designated PnFL-1, PnFL-2 and PnFL-3 , the size of which were 228 bp, 317 bp and 272 bp, respectively. A search for sequences similar to the decuced amino acid sequences was conducted using GenBank and EMBL database ; seequence encoded by PnFL-1 had 29% identity with the clone of Arabidopsis thaliana similiar to reverse trascriptase (Genbank Acc. N0.3047086), PnFL-2 shared 50% identity with hydroxiyproline-rich glycoprotein of Glycine max(GenBank Acc. No.347455), and PnFL-3 had 46% identity with TAMU 4. Thaliana genomic clone T23E16 (GenBank Acc. No.B67574). None of them was known gene in the plant system up to date, implying that the fragments may comprise parts of genes which are associated with the floral induction in Pharbitis nil.