关于安卓健的学术论文安卓健对乳腺癌细胞的诱导作用.docx

1、关于安卓健的学术论文安卓健对乳腺癌细胞的诱导作用论文标题：安卓健对乳腺癌细胞的诱导作用论文摘要：乳腺癌是全世界最流行的妇科癌症，同时乳腺癌也是全球第二大（除肺癌）造成妇女死亡的癌症。本研究，我们通过老鼠实验证实萃取自牛樟芝的环乙烯酮化合物安卓健对乳腺癌细胞具有明显的抑制其成长，促使其凋亡的作用。我们将80只雌性大白鼠分为五组，分别为AC低 剂量组（喂食0.25g牛樟芝菌丝体每千克体重）、AC高剂量组（喂食0.5g牛樟芝菌丝体每千克体重）、安卓健低剂量组（喂食15mg安卓健每千克体重）、安卓健高剂量组（喂食30mg安安卓健每千克体重）、紫杉醇组（喂食5mg紫杉醇每千克体重）。实验结果显示，无论是

2、牛樟芝组、安卓健组还是紫杉醇组，小于10mm的乳腺肿瘤均显著地缩小。但是，大于10mm的乳腺肿瘤只在安卓健组观察到显著缩小。同时发现，高剂量的牛樟芝组比低剂量的牛樟芝组，乳腺肿瘤的缩小程度更显著，并且在高剂量的牛樟芝组，小于10mm的乳腺肿瘤在17周内没有出现复发现象。英文版本论文全文如下：Name：Antrodia camphorate extract and its purified compound-Antroquinonol orally treats N- methyl -N-nitrosourea -induced breast cancer Summary: Antrodia c

3、amphorate possesses anti -breast cancer activity The Golden Biotechnology Corp. camphorata initiated by the company, many experts involved in the study the mechanism for the treatment of breast cancerRelated research. Studies have shown that A. camphorata with anti-cancer activity.Li -Hsuen Chen1, W

4、ei-Chih Yang1, Gain-He Fang1, Mao-Tien Kuo2, Wu -Che Wen2, Peini Chen3, Ching-Feng Weng1 * 1Department of Life Science and the Institute of Biotechnology, National DongHwa University, Hualien 974, Taiwan . 2 Golden Biotechnology Corp., Danshuei Township, Taipei County 251, Taiwan. 3Department of Rad

5、iology, Taipei Veterans General Hospital, Taipei 112, Taiwan. * To whom correspondence and reprint requests should be made Running title: Antrodia camphorate possesses anti -breast cancer activity _ * Corresponding author. Tel.: +886 3 8633637; fax: +886 3 8 630255. E-mail address: cfwengmail.ndhu.e

6、du.tw (Ching- Feng Weng). Abstract Breast cancer is the most prevalent cancer in women worldwide, excluding non-melanoma skin cancer, and is the second leading cause of cancer deaths in women (following lung cancer). In this study, Antrodia camphorate mycelium with medium extract (AC) and its purifi

7、ed compound - Antroquinonol were applied to treat in situ breast cancer in an N -methyl-N -nitrosourea (MNU)-induced rat model to evaluat e the therapeutic potential of A. camphorate. Eighty female Sprague -Dawley rats (21 days old) were fed ad libitum with standard rat chow and were injected intrap

8、eritoneally three times with 25 mg MNU /kg Bwt. Rats were palpated every week to check the size of the tumors and the tum or dimensions were measured using a caliper. Breast cancer rats were divided into three groups. Four groups were orally treated with (1) low (0.25 g/kg Bwt) and (2) high (0.5 g/k

9、g Bwt) doses of AC; (3) low (15 m g/kg Bwt) and (4) high (30 mg/kg Bwt) doses o f Antroquinonol for 5 weeks, while the other group was given 5 mg Paclitaxel /kg Bwt intraperitoneally five times . The results showed that rat breast tumors smaller than 10 mm were markedly diminished in the Paclitaxel

10、-, Antroquinonol - and AC-treated groups, however, tumors larger than 10 mm were not reduce d among the Paclitaxel and AC groups. Examinations of the tumors smaller than 10 mm showed that the high dose AC group had more tumor shrinkage compared to the low dose group. Moreover, less than 10 mm in dim

11、ension after high dose AC treatment exhibited no recurrence within 17 weeks. In conclusion , Antrodia camphorate and its purified compound -Antroquinonol possesses anti -tumor activity for treating the early stage s of breast cancer. Keywords: Antrodia camphorate, Antroquinonol , Paclitaxel, N -meth

12、yl-N -nitrosourea, in situ breast cancerIntroduction Edible mushrooms including Ganoderma lucidum (Reishi), Lentinula . edodes (shiitake), Antrodia camphorate (niu -chang-chih, Chang - chih) have been widely used as a medicinal fungus. G. lucidum has also been used to suppress cell adhesion and cell

13、 migration of highly invasive breast and prostate cancer cells, suggesting its potency to reduce tumor invasiveness (reviewed in Thyagarajan et al., 2007 ). Recently, one repo rt demonstrated that combinations of green tea with G. lucidum extracts show potential for the suppression of growth and inv

14、asiveness of metastatic breast cancer (MDA-MB-231) cells ( Thyagarajan et al., 2007 ). Moreover, structurally related lanostane -type triterpenes, including ganoderic acid A, F and H (GA -A, GA- F, GA-H), have been identified in an oriental medicinal mushroom G. lucidum. GA- A and GA-H exert their b

15、iological effects through inhibition of the transcription factors AP-1 and NF-B, resulting in the down-regulation of expression of Cdk4 and suppression of secretion of uPA, respectively ( Jiang et al., 2008). Lentinula edodes (shiitake) aqueous extracts have demonstrated direct inhibition of the pro

16、 liferation of human breast cancer cells in vitro and immuno-stimulatory properties in terms of mitogenic and co -mitogenic activity in vitro ( Israilides et al., 2008 ). A. camphorata, one of the extensively used medicinal mushrooms, is well known in Taiwan as a traditional Chinese medicine and has

17、 recently been identified as belonging to the new genus of the Antrodia species ( Taiwanofungus). Its fruiting body has been utilized in traditional Chinese medicine for the treatment of food and drug intoxication, diarrhea, abdominal pain, hypertension, skin itches, liver cancer, anti -inflammatory

18、 , anti -oxidati on, induced vasorelaxation, decreased hepatitis B virus, and h ypotriglyceride (reviewed in Suk et al., 2008 ). However, the A. camphorata fruiting bod ies are ve ry rare and expensive, and little has been reported about its biological activity, especially in treating the breast can

19、cer. Breast cancer is the most prevalent cancer in women worldwide, excluding non-melanoma skin cancer, and is the second leading cause of cancer death in women (following lung cancer). Breast cancer remains a global public health problem with some 1.1 million women newly diagnosed with breast cance

20、r in 2002 (Ferlay et al., 2002), with approximately 185,000 new cases and over 40,000 deaths estimated in the USA in 2008 ( Jemal et al., 2008). Therefore, developing a suitable animal model for testing the anti -breast cancer potency of A. camphorata becomes a high priority. N -methyl-N -nitrosoure

21、a (MNU) is an alkylating agent that methylates DNA bases at nucleophilic sites (generating N7 and N3 alkylpurines). The primary mutagenic lesion is O6 methylguanine. MNU-induced mammary tumorigenesis in rats is one of the most broadly used rodent mammary tumorigenesis models for studying human breas

22、t cancer (Russo et al., 1990; Gould, 1995; Nandi et al., 1995 ). MNU - induced rat mammary tumorigenesis and human breast cancer development share many similarities in hormone dependency, pathogenesis, histological classification, and immunocytochemical markers. Based on these similarities, MNU -ind

23、uced rat mammary tumorigenesis appears to be a useful experimental model for the study of human breast cancer development (Thordarson et al., 2001 ). Antroquinonol (1), an ubiquinone derivative, is isolated from t he solid-state fermented mycelium of A. camphorate , a parasitic fungus indigenous to

24、Taiwan. The structure of compound 1 i s elucidated by the analysis of their spectroscopic data . It contains anti-tumor activity ( Lee et al., 2007). Recently, A. camphorate extract has been proven for the treatment of systemic lupus erythematosus (SLE) in SLE-prone NZB/W F1 mice (Chang et al., 2008

25、). The aim of this study was to produce breast cancer in rats to evaluate the anti -tumor activity of A. camphorate extract and its purified compound- Antroquinonol for treating in situ breast cancer. Materials and methods Chemicals N -methyl-N -nitrosourea (MNU, N4766) and Paclitaxel ( TaxolTM, T71

26、91) were purchased from Sigma - aldrich (St. Louis, CA, USA). The pure Antroquinonol compo und and A. camphorata extract (ACE) were obtained from Golden Biot echnology Company, Taiwan . Preparation of A. camphorata extract The A. camphorata (strain no. 486bp) used in this study was authenticated by

27、Dr Tzean Shang-Shong, Department of Plant Pathology and Microbiology, National Taiwan University, Taiwan. The mycelia of A. camphorata were cultured in 1000 mL of growth medium containing 0.1 g NaCl, 10 g peptone, 2 g yeast extract, 10 g agar and 10 g cereal mixture (rice, wheat or corn), pH 7.5 at

28、25oC for 12 14 weeks. After cultivation, 500 g of lyophilized A. camphorata mycelium was extracted with 2500 mL of n-hexane for 6 h. The n-hexane fraction, defined as A. camphorate extract (GD -66, product name and GD -AIDT7, batch name), was concentrated to 2030 mL via vacuum evaporation . A. camph

29、orate extract has been approved as a health food by Taiwan FDA (Healthy Food No. A00124). The purified procedure of Antroquinonol was described in our previous study ( Lee et al., 2007). Briefly, Antroquinonol was purif ied from A. camphorate extract by silica gel chromatography (column=45 cm, 5 cm

30、i.d.) (ASTM silica gel, Merck Co., Germany) and eluted with n-hexane:ethyl acetate (10 : 3 v/v). The resulting eluate was further subjected to size exclusion chromatography using Sephadex LH20 column (70 cm long, 5 cm i.d.) (AB gel, GE Healthcare Bio-Science, USA). The antroquinonol (98% purity) was

31、 eluted with 95% ethanol; the A. camphorate extract fraction was determined to have 0.01% antroquinonol . Animals and treatments Twenty - one-day-old fem ale Sprague-Dawley rats were obtained from the BioLASCO Taiwan Co., Ltd (Taipei, Taiwan) and were kept in a temperature -controlled environment (2

32、 3oC ) with 70% relative humidity under a 12-h light/dark cycle. The animal experiments were pe rformed according to the Guide for the Care and Use of Laboratory Animals of National Dong-Hwa University. Rats were fed standard rat chow (LabDietTM -500l Rodent Diet; TestDiet, Richmond, IN, USA). After one week adaptation, all rats re