Betulin Attenuates TGF-ß1- and PGE2-Mediated Inhibition of NK Cell Activity to Suppress Tumor Progression and Metastasis in Mice.

Transforming growth factor (TGF)-ß1 and prostaglandin E2 (PGE2) are humoral factors critically involved in the induction of immunosuppression in the microenvironment of various types of tumors, including melanoma. In this study, we identified a natural compound that attenuated TGF-ß1- and PGE2-induced immunosuppression and examined its effect on B16 melanoma growth in mice. By screening 502 natural compounds for attenuating activity against TGF-ß1- or PGE2-induced suppression of cytolysis in poly(I:C)-stimulated murine splenocytes, we found that betulin was the most potent compound. Betulin also reduced TGF-ß1- and PGE2-induced downregulation of perforin and granzyme B mRNA expression and cell surface expression of NKG2D and CD69 in natural killer (NK) cells. Cell depletion and coculture experiments showed that NK cells, dendritic cells, B cells, and T cells were necessary for the attenuating effects of betulin. Structure-activity relationship analysis revealed that two hydroxyl groups at positions C3 and C28 of betulin, their cis-configuration, and methyl group at C30 played crucial roles in its attenuating activity. In a subcutaneous implantation model of B16 melanoma in mice, intratumor administration of betulin and LY2157299, a TGF-ß1 type I receptor kinase inhibitor, significantly retarded the growth of B16 melanoma. Notably, betulin increased significantly the number of CD69 positive NK cells in tumor sites at early stages of post-tumor cell injection. Our data suggest that betulin inhibits the growth of B16 melanoma by enhancing NK cell activity through attenuating the immunosuppressive tumor microenvironment.

Stabilizers for Osmium Isotopes in Microwave-Irradiated Acid Digestion and Inductively Coupled Plasma Mass Spectrometry Analysis.

Osmium is defined in the international council for harmonization (ICH-Q3D) guidelines as an element whose concentration can be determined by validated methods including microwave-assisted nitric acid digestion and inductively coupled plasma mass spectrometry. However, microwave digestion using nitric acid is known to result in osmium recoveries higher than the theoretical values in spiked tests because of the formation of highly volatile osmium tetroxide in an oxidation reaction. To stabilize osmium, the addition of thiourea as a complexing agent has been tested and proved its utility. It remains unclear whether other compounds can prevent the over-recovery of osmium. In this study, we investigated four compounds, thiourea, ascorbic acid, sodium sulfite, and potassium metabisulfite, that could reduce the overestimation of osmium isotopes. The minimum amounts of thiourea, ascorbic acid, sodium sulfite, and potassium metabisulfite required to stabilize 10 ng/mL osmium in blank matrix were 1.0, 1.0, 2.5, and 2.5 g/L, respectively. The relative standard deviations obtained from 12 analyses for each stabilization solution were less than 3.3% in thiourea, 12.7% in ascorbic acid, 9.0% in sodium sulfite, and 10.6% in potassium metabisulfite. The stabilization solutions were investigated in a digested tablet matrix and were found to be effective. The impact of adding stabilization solutions on the determination of all ICH-Q3D element concentrations was also evaluated. As stabilization solutions had a small or significant impact on the determination of some elements, it was concluded that osmium determination should be conducted independently.

A Liquid Chromatography-Mass Spectrometry Method to Study the Interaction between Membrane Proteins and Low-Molecular-Weight Compound Mixtures.

Molecular interaction analysis is an essential technique for the study of biomolecular functions and the development of new drugs. Most current methods generally require manipulation to immobilize or label molecules, and require advance identification of at least one of the two molecules in the reaction. In this study, we succeeded in detecting the interaction of low-molecular-weight (LMW) compounds with a membrane protein mixture derived from cultured cells expressing target membrane proteins by using the size exclusion chromatography-mass spectrometry (SEC-MS) method under the condition of 0.001% lauryl maltose neopentyl glycol as detergent and atmospheric pressure chemical ionization. This method allowed us to analyze the interaction of a mixture of medicinal herbal ingredients with a mixture of membrane proteins to identify the two interacting ingredients. As it does not require specialized equipment (e.g., a two-dimensional liquid chromatography system), this SEC-MS method enables the analysis of interactions between LMW compounds and relatively high-expressed membrane proteins without immobilization or derivatization of the molecules.

Bidirectional crosstalk between neutrophils and adipocytes promotes adipose tissue inflammation.

Chronic activation of the IL-1ß system in adipose tissue on metabolic disorders is well demonstrated. However, a mechanism for its expression and activation in the tissue has remained unexplored. Here, we demonstrate that IL-1ß transcript was enriched in neutrophils of white adipose tissue (WAT) from lean mice. Mechanistically, the interaction of neutrophils with adipocytes induced IL-1ß expression via NF-κB pathway. Lipolysis of adipocytes accumulated neutrophils prior to macrophages in WAT and produced high levels of IL-1ß via an inflammasome pathway. Leukotriene B4 (LTB4) production in WAT also contributed to neutrophil accumulation. Furthermore, an LTB4-inflammasome axis contributed to the expression of chemotactic molecules involved in high-fat diet-induced macrophage infiltration into WAT. We have identified previously unappreciated roles for neutrophils in the development of adipose tissue inflammation: robust IL-1ß production and infiltration of macrophages to initiate chronic inflammation.-Watanabe, Y., Nagai, Y., Honda, H., Okamoto, N., Yanagibashi, T., Ogasawara, M., Yamamoto, S., Imamura, R., Takasaki, I., Hara, H., Sasahara, M., Arita, M., Hida, S., Taniguchi, S., Suda, T., Takatsu, K. Bidirectional crosstalk between neutrophils and adipocytes promotes adipose tissue inflammation.

Oncologic outcomes for open and laparoscopic radical nephroureterectomy in patients with upper tract urothelial carcinoma.

BACKGROUND: Oncologic benefits of laparoscopic radical nephroureterectomy (LNU) are unclear. We aimed to evaluate the impact of surgical approach for radical nephroureterectomy on oncologic outcomes in patients with locally advanced upper tract urothelial carcinoma (UTUC). METHODS: Of 426 patients who underwent radical nephroureterectomy at five medical centers between February 1995 and February 2017, we retrospectively investigated oncological outcomes in 229 with locally advanced UTUC (stages cT3-4 and/or cN+). The surgical approach was classified as open nephroureterectomy (ONU) or LNU, and oncologic outcomes, including intravesical recurrence-free survival (RFS), visceral RFS, cancer-specific survival (CSS), and overall survival (OS), were compared between the groups. The inverse probability of treatment weighting (IPTW)-adjusted Cox-regression analyses was performed to evaluate the impact of LNU on the prognosis. RESULTS: Of the 229 patients, 48 (21%) underwent LNU. There were significant differences in patient backgrounds, including preoperative renal function, lymph-node involvement, lymphovascular invasion, and surgical margins, between the groups. Before the background adjustment, intravesical RFS, visceral RFS, CSS, and OS were significantly inferior in the ONU group than in the LNU group. However, in the IPTW-adjusted Cox-regression analysis, no significant differences were observed in intravesical RFS (hazard ratio [HR], 0.65; P = 0.476), visceral RFS (HR, 0.46; P = 0.109), CSS (HR, 0.48; P = 0.233), and OS (HR, 0.40; P = 0.147). CONCLUSION: Surgical approaches were not independently associated with prognosis in patients with locally advanced UTUC.

The utility of neoadjuvant gemcitabine plus carboplatin followed by immediate radical cystectomy in patients with muscle-invasive bladder cancer who are ineligible for cisplatin-based chemotherapy.

BACKGROUND: There are currently no data supporting carboplatin-based regimens in muscle-invasive bladder cancer patients who are unsuitable for neoadjuvant cisplatin treatment. The aim of this study was to investigate the potential benefit of carboplatin-based regimens in patients with muscle-invasive bladder cancer who were unsuitable for cisplatin. METHODS: We focused on 171 patients ineligible for cisplatin, including 98 patients who received neoadjuvant gemcitabine and carboplatin (GCarbo) followed by immediate radical cystectomy (GCarbo cohort) and 73 patients who underwent radical cystectomy (RC alone cohort) at Hiroaki University or Aomori Prefectural Hospital. In the neoadjuvant GCarbo cohort, patients underwent two 21-day cycles of GCarbo the two courses of neoadjuvant chemotherapy were followed by radical cystectomy at an interval of 1 month. RESULTS: Of the enrolled patients, 165 (95.3 %) had renal impairment. The median age of the enrolled patients was 71 years, and the median follow-up period was 63 months. The 5-year overall survival rates for the GCarbo and the RC-alone cohorts were 79.5 and 53.8 %, respectively (P < 0.001), while the 5-year disease-free survival rates were 75.5 and 55.4 %, respectively (P = 0.013). Surgical specimens of 16 (16.3 %) patients in the GCarbo cohort indicated stage pT0 disease. The rate of positive surgical margins in the RC-alone cohort was significantly higher than that in the GCarbo cohort (P < 0.001). CONCLUSIONS: In this study, the oncological outcomes were significantly improved in cisplatin-unfit patients with muscle-invasive bladder cancer who received neoadjuvant GCarbo chemotherapy, compared with those in patients who underwent RC alone. The standard of care for muscle-invasive bladder cancer, especially for patients who are unfit for cisplatin, may include neoadjuvant carboplatin chemotherapy followed by radical cystectomy.

Identification of innate IL-5-producing cells and their role in lung eosinophil regulation and antitumor immunity.

IL-5 is involved in a number of immune responses such as helminth infection and allergy. IL-5 also plays roles in innate immunity by maintaining B-1 B cells and mucosal IgA production. However, the identity of IL-5-producing cells has not been unambiguously characterized. In this report, we describe the generation of an IL-5 reporter mouse and identify IL-5-producing non-T lymphoid cells that reside in the intestine, peritoneal cavity, and lungs in naive mice. They share many characteristics with natural helper cells, nuocytes, and Ih2 cells, including surface Ags and responsiveness to cytokines. However, these phenotypes do not completely overlap with any particular one of these cell types. Innate non-T IL-5-producing cells localized most abundantly in the lung and proliferated and upregulated IL-5 production in response to IL-25 and IL-33. IL-33 was more effective than IL-25. These cells contribute to maintaining sufficient numbers of lung eosinophils and are important for eosinophil recruitment mediated by IL-25 and IL-33. Given that eosinophils are shown to possess antitumor activity, we studied lung tumor metastasis and showed that innate IL-5-producing cells were increased in response to tumor invasion, and their regulation of eosinophils is critical to suppress tumor metastasis. Genetic blockade or neutralization of IL-5 impaired eosinophil recruitment into the lung and resulted in increased tumor metastasis. Conversely, exogenous IL-5 treatment resulted in suppressed tumor metastasis and augmented eosinophil infiltration. These newly identified innate IL-5-producing cells thus play a role in tumor surveillance through lung eosinophils and may contribute to development of novel immunotherapies for cancer.

Long-term disease control after upfront chemotherapy and surgery in a patient with primary prostate leiomyosarcoma.

INTRODUCTION: Prostate leiomyosarcoma is a rare, aggressive neoplasm. CASE PRESENTATION: A 52-year-old man presented with worsening frequent micturition and painful urination. Rectal examination revealed a significantly enlarged prostate. Magnetic resonance imaging showed a large prostate tumor with urinary bladder and bilateral seminal vesicle invasion. A prostate biopsy revealed diffuse proliferation of pleomorphic atypical cells. Immunohistochemistry confirmed the diagnosis of prostrate leiomyosarcoma. The patient received three cycles of the mesna, doxorubicin, ifosfamide, and dacarbazine regime (mesna 6000 mg/m2, doxorubicin 60 mg/m2, ifosfamide 7500 mg/m2, and dacarbazine 900 mg/m2) at 4-week intervals. The tumor shrank by 28% and exhibited necrotic changes. He underwent total pelvic exenteration with en bloc resection of the prostate, bladder, rectum, and anus. Pathological surgical margin was negative. The patient is alive with no disease at 5 years postoperatively. CONCLUSION: Neoadjuvant chemotherapy and surgical resection are essential to achieve a long-term survival of patients with localized prostate leiomyosarcoma.

[Chemo-endocrine therapy with low-dose cisplatin, UFT, and dexamethasone for hormone-refractory prostate cancer patients].

Since September 2005, twenty-two patients with hormone-refractory prostate cancer (aged 55-81 years) were treated with LH-RH agonist and low-dose cisplatin, UFT, and dexamethasone after proving resistant to estramustine phosphate therapy. The regimen of this therapy consists of 5 mg/body of cisplatin intravenously once a week, 300 mg/day of UFT and 1 mg/day of dexamethasone orally, every day. All patients suffered from clinical progression such as local recurrence in 11 patients who had already received radiation therapy, lymph node metastasis in 7 patients, and bone metastasis in 15 patients. Initial PSA value ranged from 1.7 ng/mL to 215.1 ng/mL. The PSA response rate, which decreased more than 50% in PSA values was 72. 7% (16/22). The follow-up term ranged from 2 to 43 months, and nine patients died of cancer progression. The median time to progression was 11 months, and median overall survival was 19 months. There were no severe adverse effects, and stoppages of the therapy for 13 patients were all due to disease progression. Following this therapy, 9 patients received best supportive care and 4 patients received docetaxel chemotherapy. We considered this therapy to be effective for patients with hormone-refractory prostate cancer because it maintained their good QOL.

Risk-stratified surveillance protocol improves cost-effectiveness after radical nephroureterectomy in patients with upper tract urothelial carcinoma.

OBJECTIVES: To develop a surveillance protocol with improved cost-effectiveness after radical nephroureterectomy (RNU), as the cost-effectiveness of oncological surveillance after RNU remains unclear. RESULTS: Of 426 patients, 109 (26%) and 113 (27%) experienced visceral and intravesical recurrences, respectively. The pathology-based protocol found significant differences in recurrence-free survival in the visceral recurrence but not in the intravesical recurrence. The medical costs per visceral recurrence detected were high, especially in normal-risk (≤ pT2N0, LVI-, SM-) patients. We developed a risk score associated with visceral recurrence using Cox regression analysis. The risk score-based protocol was significantly more cost-effective than the pathology-based protocol. Estimated cost differences reached $747,929 per recurrence detected, a suggested 55% reduction. MATERIALS AND METHODS: We retrospectively evaluated 426 patients with RNU for upper tract urothelial carcinoma (UTUC) without distant metastasis at 4 hospitals. Patients with routine oncological follow-up were stratified into normal-, high- and very high-risk groups according to a pathology-based protocol utilizing pathological stage, lymphovascular invasion (LVI) and surgical margin (SM). Cost-effectiveness of the pathology-based protocol was evaluated, and a risk score-based protocol was developed to optimize cost-effectiveness. Risk scores were calculated by summing up risk factors independently associated with recurrence-free survival. Patients were stratified by low-, intermediate- and high-risk score. Estimated cost per recurrence detected by pathology-based and risk score-based protocols was compared. CONCLUSIONS: A risk score-stratified surveillance protocol has the potential to reduce over investigation during follow-up, making surveillance more cost-effective.

Functional Characterization of Epitheaflagallin 3-O-Gallate Generated in Laccase-Treated Green Tea Extracts in the Presence of Gallic Acid.

Epitheaflagallin (ETFG) and epitheaflagallin 3-O-gallate (ETFGg) are minor polyphenols in black tea extract that are enzymatically synthesized from epigallocatechin (EGC) and epigallocatechin gallate (EGCg), respectively, in green tea extract via laccase oxidation in the presence of gallic acid. The constituents of laccase-treated green tea extract in the presence of gallic acid are thus quite different from those of nonlaccase-treated green tea extract: EGC and EGCg are present in lower concentrations, and ETFG and ETFGg are present in higher concentrations. Additionally, laccase-treated green tea extract contains further polymerized catechin derivatives, comparable with naturally fermented teas such as oolong tea and black tea. We found that ETFGg and laccase-treated green tea extracts exhibit versatile physiological functions in vivo and in vitro, including antioxidative activity, pancreatic lipase inhibition, Streptococcus sorbinus glycosyltransferase inhibition, and an inhibiting effect on the activity of matrix metalloprotease-1 and -3 and their synthesis by human gingival fibroblasts. We confirmed that these inhibitory effects of ETFGg in vitro match well with the results obtained by docking simulations of the compounds with their target enzymes or noncatalytic protein. Thus, ETFGg and laccase-treated green tea extracts containing ETFGg are promising functional food materials with potential antiobesity and antiperiodontal disease activities.

[The effects of the antagonists of muscarinic acetylcholine receptor subtypes in rat brain on urinary bladder contraction].

BACKGROUND: Pontine micuturition center and storage center are controlled by upper brain. Pharmacological and molecular biological data indicate the distribution of muscarinic receptor subtypes in rat brain, however, the effect of central muscarinic cholinergic mechanisms on the bladder activity has not been evaluated. We investigated the diversity of effect of each subtype in rat brain. METHODS: The muscarinic agents such as muscarine (non-specific agonist), atropine (non-specific antagonist), pirenzepine (M1 receptor antagonist), AF-DX116 (M2 receptor antagonist) and 4-DAMP (M3 receptor antagonist) were administrated with intracerebroventricular injection (I.C.V. group) or intravenous injection (i.v. group). The drug effects on rhythmic bladder contraction were investigated. RESULTS: In I.C.V. group, the bladder contraction pressure were reduced by atropine, pirenzepine and 4-DAMP. The contraction time were shortened by pirenzepine and 4-DAMP but extended by AF-DX116. The contraction frequency was decreased by AF-DX116. Whereas AF-DX116 in i.v. group reduced the contraction pressure but did not cause any changes of the contraction time and the frequency. Other agents in i.v. group had same tendency with those of I.C.V. group except for intensity. CONCLUSION: That is to say AF-DX116 in I.C.V. group facilitate both micuturition center and storage center. In other words, these data indicate M2 receptor in rat brain inhibits both micurition and storage center. In addition, the effect of AF-DX116 in brain suggests that the development of new drug which proceed to central neuron system might provide a new treatment of neurogenic bladder.

Primary leiomyosarcoma of a horseshoe kidney in a woman with Turner syndrome: a case report.

BACKGROUND: Turner syndrome is characterized by complete or partial X-chromosome monosomy and has various clinical features, including horseshoe kidney. Leiomyosarcoma is an extremely rare tumor that accounts for only 0.1% of all invasive renal tumors. CASE PRESENTATION: A 50-year-old Japanese woman presented at a community hospital with a chief complaint of abdominal pain. Computed tomography revealed a horseshoe kidney with a hypovascular tumor (size, 9 × 7 cm) showing calcification in the upper pole of the right kidney. Open right heminephrectomy and division of the isthmus were performed. Histological examination revealed alternating fascicles of spindle cells with blunt ended non-tapering nuclei and eosinophilic cytoplasm. The tumor had high mitotic activity with a mitotic count of 8 mitoses/10 high-power fields. On the basis of these findings, we diagnosed the patient as having leiomyosarcoma. CONCLUSION: Primary leiomyosarcoma of the horseshoe kidney in a patient with Turner syndrome is a very rare occurrence.

Role of fatty acid chain length on the induction of apoptosis by newly synthesized catechin derivatives.

The catechins, a family of polyphenols found in tea, can evoke various responses, including apoptosis. In this study we investigated whether the chemical modification of (-)-epigallocatechin gallate (EGCG) could enhance its apoptosis activity. We found that one of the catechin conjugated with capric acid [(2R,3S)-3',4',5,7-tetrahydroxyflavan-3-yl decanoate; catechin-C10] was most potent to induce apoptosis in U937 cells. C10 treatment resulted in a significant increase in reactive oxygen species (ROS) formation, mitochondrial membrane potential (MMP) loss, cytochrome c release caspase-9 and caspase-3 activation. In addition to this C10 also activated extrinsic pathway significantly as evident by time-dependent increase in Fas expression and caspase-8 activity. C10 mediated cleavage of Bid may be an important event for cross talk between intrinsic and extrinsic signaling. Moreover, pre-treatment of cells with anti-oxidant N-acetyl-L-cysteine (NAC) significantly prevented C10-induced apoptosis but did not protect MMP loss. Treatment of cells with pan-caspase inhibitor significantly inhibited apoptosis indicating that caspases are playing key role. In addition to this C10 was found to induce apoptosis in human colon cancer (HCT116) cells while it showed resistance to human keratinocytes (HaCat). In short our results showed that the optimal fatty acid side chain length is required for the apoptosis inducing activity of catechin derivatives in U937 cells.

Differential effects of antioxidants on the in vitro invasion, growth and lung metastasis of murine colon cancer cells.

We conducted a comparative study of 20 antioxidants including antioxidative vitamins and polyphenols to examine their inhibitory activities against the in vitro invasion, growth and experimental lung metastasis of murine colon 26-L5 carcinoma cells. Among the compounds tested, epigallocatechin gallate (EGCG), gallocatechin gallate and genistein exhibited significant reductions at 77%, 46% and 44% in tumor metastasis by an intraperitoneal administration for 5 d beginning at 3 d before tumor inoculation, respectively. Quercetin also showed a slight but not statistically significant inhibition. Alpha-tocopherol, beta-carotene, ascorbic acid and 2 EGCG-related compounds of epicatechin gallate and epigallocatechin had no effect. EGCG also inhibited tumor metastasis dose-dependently with 98% suppression at 2 micromol; and an almost equivalent inhibition was also produced by only pre-administration of EGCG at the same dose before tumor inoculation. EGCG significantly inhibited tumor cell invasion and proliferation, but its inhibition of these activities was much less effective than that of other compounds which did not show any antimetastatic effect. No statistically significant relationship was observed between the radical scavenging activities of the test compounds and their rates of inhibition of tumor metastasis. The antimetastatic mechanism of EGCG thus seems to be independent of its inhibition of tumor invasion and growth, as well as its radical scavenging activity. Our results suggest that EGCG is potentially beneficial for tumor metastasis inhibition.

New plastoquinones isolated from the brown alga, Sargassum micracanthum.

Four new plastoquinones were isolated from the methanolic extract of the brown alga, Sargassum micracanthum. Their structures were elucidated based on spectroscopic analysis and chemical conversions from 2-geranylgeranyl-6-methyl-1,4-benzohydroquinone. These plastoquinones exhibited significant antioxidant activities such as an inhibitory effect on lipid peroxidation and a radical scavenging effect on 1,1-diphenyl-2-picrylhydrazyl (DPPH). Some of the new plastoquiones showed cytotoxic activity against cancer cell line.

Inhibition by evodiamine of hepatocyte growth factor-induced invasion and migration of tumor cells.

Tumor cell motility plays a crucial role in the establishment of tumor metastasis and is affected by a variety of host-derived factors during the event. Hepatocyte growth factor (HGF) is one of these factors and stimulates tumor cell migration remarkably. We previously reported that evodiamine has a marked inhibitory activity on tumor cell invasion and migration in vitro. In this study, the effects of evodiamine on HGF-induced invasion and migration of tumor cell lines, colon 26-L5 carcinoma, B16-F10 melanoma and Lewis lung carcinoma (LLC) were examined. HGF promoted invasive activity of tumor cell lines with maximal induction of 1.8 times at 30 ng/ml for colon 26-L5 and LLC cells, and 2.0 times at 10 ng/ml for B16-F10 cells. Evodiamine inhibited the HGF-stimulated tumor cell invasion and migration in a concentration-dependent manner, and achieved complete suppression at 30 microM in all of the cell lines tested. When tumor cells were seeded on fibronectin-coated plates with evodiamine, their spreading on the plate was obviously inhibited, while their adhesiveness to fibronectin was unaffected. Evodiamine showed a marginal effect on tumor cell growth in a 24-h incubation, although it exhibited a marked inhibition in an over 48-h incubation. These results suggest that evodiamine suppressed HGF-stimulated invasion and migration of tumor cells partly through inhibition of cell spreading.

Anti-invasive and metastatic activities of evodiamine.

We have recently reported that evodiamine can suppress in vitro invasion and lung metastasis by colon 26-L5 carcinoma cells. To extend our study, we examine here the anti-invasive and metastatic effects of evodiamine on Lewis lung carcinoma (LLC) and B16-F10 melanoma in addition to colon 26-L5 carcinoma. Critical structures of evodiamine for the activities were also evaluated by comparison with compounds possessing structures similar to that of evodiamine. Evodiamine concentration-dependently inhibited the invasion of B16-F10, LLC and colon 26-L5 cells with IC(50) values of 2.4 micro M, 4.8 micro M and 3.7 micro M, respectively. Pre-treatment of colon 26-L5 cells with evodiamine before inoculation into mice caused significant suppression of the liver metastasis as well as the lung metastasis. Lung metastasis by LLC is also inhibited significantly by pre-exposure to evodiamine. When the anti-migratory activity of evodiamine was compared with that of evodiamine-like compounds, rutaecarpine lacking a methyl group at N-14 and a hydrogen at C-13 b exhibited much less effect than evodiamine. In addition, reserpine, having beta-configurated hydrogen at C-13 b, inhibited tumor cell migration more potently than yohimbine, having alpha-configurated hydrogen at the same position. These results suggest that evodiamine may be useful as a leading compound for agents in tumor metastasis therapy. Also, the presence of a methyl group at N-14 and the configuration of hydrogen at C-13 b may be responsible for the inhibitory activities of evodiamine.