臺灣博碩士論文加值系統

超臨界萃取技術具有低溫萃取、低黏性及高擴散能力之特性，能將對熱不穩
定之物質完整的萃取，且保留其活性成分，故能將樟芝所含的活性物質萃取。樟
芝又名牛樟芝或牛樟菇學名為Antrodia camphorata，是台灣特有真菌類，其生物
活性具有抗腫瘤， 抗發炎，抗氧化等作用。因此本研究主要以固態培養
樟芝菌絲體在不同壓力下之超臨界樟芝萃取進行生物活性探討。結果顯
示經由固態培養樟芝水萃取物及不同壓力下超臨界樟芝萃取粉末之水萃取物處
理後隨著濃度增加而對乳癌細胞(MCF-7、MDA-MB 231)增生之抑制率增加，能夠
顯著抑制乳癌細胞( MCF-7、MDA-MB 231)的生長，更進一步利用流式細胞儀於細
胞凋亡上發現，隨著濃度增加促使細胞走向凋亡的現象，此外，透過RT-PCR 發現，
在細胞凋亡路徑中，經由固態培養樟芝水萃取物及不同壓力下超臨界樟芝萃取粉
末之水萃取物處理後，Bcl-2 mRNA 的表現量隨著濃度增加明顯被抑制，而對Bax
mRNA 表現量明顯升高；於西方墨點法試驗中也發現會影響乳癌細胞內Bcl-2 、
Bax 蛋白質之表現量，導致細胞凋亡現象。在抑制轉移試驗中發現固態培養樟芝菌
絲體之水萃物以及經由不同壓力下超臨界樟芝萃取粉末之水萃取物，處理HT-1080
分泌基質金屬蛋白酶MMP-2 及MMP-9，明顯抑制效果，此外在RT-PCR 試驗中
也發現，經由樟芝水萃及經由不同壓力下超臨界樟芝萃取粉末之水萃取物處理
後，細胞MMP-2 及MMP-9 mRNA 表現也受到抑制。在DCFH-DA 抗氧化試驗中，
經由流式細胞儀發現固態培養樟芝菌絲體之水萃物及經由不同壓力下超臨界樟芝
萃取粉末之水萃取物，處理CHO-K1 細胞，能夠減少細胞內ROS 含量，具有抗氧
化活性。在提升免疫試驗中發現，利用固態培養樟芝菌絲體之水萃物及經由不同
壓力下超臨界樟芝萃取粉末之水萃取物，對Raw 264.7 巨噬細胞刺激後能促使細
胞分泌細胞激素而抑制乳癌細胞(MCF-7、MDA-MB 231 )增生。綜合以上結果證實
固態培養樟芝菌絲體之水萃物及經由不同壓力下超臨界樟芝萃取粉末之水萃取物確實有抗腫瘤、抑制轉移以及抗氧化之生物活性，實驗結果顯示樟芝及超臨界方
面值得開發。

The supercritical extraction technology, such as with low temperature extraction,
low viscosity and high diffusion capacity, has some benefit for food industry.
Therefore, these kinds of characteristics are good for unstable substances to retain their
active ingredient. Although many papers have shown that Antrodia camphorata has
several biological activities such as anti-tumor, anti-inflammatory, and anti-oxidation
effects, there are lots of biological functions still need to be elucidated. In order to
investigate the bioactivities of A. camphorata, the biological activities of the
supercritical fluid extraction (SFE) from A. camphorata under different pressures were
evaluated during this study. The results showed that the SFF from A. camphorata can
significantly inhibit the cell growth of breast cancer cells (MCF-7 and MDA-MB-231)
due to MTT assay. Moreover, the growth-inhibition effect in breast carcinoma cells
was due to apoptosis evaluated by flow cytometer. In addition, the pathway of
apoptosis on breast carcinoma cells was further confirmed by RT-PCR assay and
werstern blot anslyasis such as the Bax and Bcl-2 expression. Our data also showed
that SFE from A. camphorata has growth-inhibitory effect in tumor cells. In
antioxidant assay, we found that water extract or SFE of mycelia from A. camphorata
can reduce CHO-K1 cell death caused by H2O2. According immunomodulatory assay,
the results showed that the water extract or SFE can modulate cytokine production in
Raw 264.7 cells. Therefore, our data suggest that water extract or SFE from A.
camphorata have anti-tumor, anti-metastasis, antioxidant activity as well as
immunomodulatoy effect.

目 錄
中文摘要 i
英文摘要 iii
第一章 文獻回顧 1
一、研究背景 1
二、樟芝( Antrodia camphorata )簡介 1
2.1 特性 1
2.2 生理活性成分 2
三、超臨界流體 (Supercritical fluid，SCF) 3
3.1 超臨界流體發展 3
3.2 超臨界流體定義 3
3.3 超臨界流體萃取( Supercritical Fluid Extraction，SFE)基本原理及應用
4
四、癌症 5
4.1 乳癌的介紹 5
4.2 乳癌的治療 6
五、雌激素與雌激素受器 7
六、癌細胞的轉移與細胞凋亡 8
6.1 腫瘤轉移 8

6.2 細胞凋亡定義 9
6.3 細胞凋亡與細胞壞死 9
6.4 細胞凋亡之訊息調控路徑 10
第二章 研究動機 12
第三章 實驗架構 13
第四章 材料方法 14
一、實驗材料 14
1.1 實驗細胞株. 14
二、實驗方法 14
2.1 樣品製備 14
2.1.1 水萃取法 14
2.1.2 超臨界萃取方法 15
2.2 細胞存活率測定( MTT assay ) 15
2.2.1 實驗步驟 15
2.3 Gelatin Zymograpgy Assay 16
2.3.1Conditioned medium 與細胞溶解液（cell lysate）的收集 16
2.3.2 蛋白質含量的測定 17
2.4 電泳分析法 17
2.4.1 方法 17

2.5 細胞凋亡檢測 18
2.5.1 實驗步驟 18
2.6 西方墨點法(Western Bolt) 18
2.7 反轉錄聚合酵素鏈鎖反應 19
( RT-PCR，reverse transcription polymerase chain reaction)
2.7.1 mRNA 萃取 19
2.7.2 反轉錄聚合酶連鎖反應(RT-PCR) 20
2.7.3 電泳分析法cDNA 分析片段 ( Agarose gel electrophoresis ) 21
2.8 抗氧化能力測試 21
2.9 提升免疫能力測試. 21
2.9.1 實驗步驟 22
2.9.2 抑制癌細胞株生長試驗 22
2.10 統計分析 22
第五章 結果 24
一、抑癌試驗 24
1.1 抑制腫瘤細胞增生試驗 24
1.2 W、SFE-250 bar 及SFE-400 bar 對乳癌細胞之細胞凋亡影響 25
1.3 W、SFE-250 bar 及SFE-400 bar 對乳癌細胞Bcl-2、Bax mRNA 表
現之影響 27

1.4 W、SFE-250 bar 及SFE-400 bar 對乳癌細胞Bcl-2、Bax 、Pro-caspase-9
蛋白質表現之影響 27
二、轉移酵素試驗 28
三、W、SFE-250 bar 及SFE-400 bar 對細胞MMP-2 與MMP-9 m RNA 表
現的影響 28
四、W、SFE-250 bar 及SFE-400 bar 之細胞抗氧化測定 29
五、W、SFE-250 bar 及SFE-400 bar 免疫能力試驗 29
第六章 討論 32
一、抑制腫瘤細胞存活試驗 32
二、細胞凋亡試驗 32
三、樟芝水萃物(W、SFE-250 bar 及SFE-400 bar)對乳癌細胞Bax、Bcl-2 表
現之影響 33
四、MMPs 活性試驗 34
五、樟芝之細胞內抗氧化測定 34
六、間接免疫與免疫能力活性試驗 34
第七章 結論 35
第八章 未來方向 37
第九章 參考文獻 39

表目錄
表一、RT-PCR 分析所使用之primer 以及試驗的條件。 20
表二、W、SFE-250 bar、SFE-400 bar 對乳癌細胞(MCF-7、MDA-MB-231)存活
之影響。 24
表三、以不同濃度之樟芝及超臨界樟芝粉末水萃物(SFE-250 bar 及SFE-400 bar)
處理MCF-7 與MDA-MB-231 細胞12 hr 後細胞凋亡之變化。 26
表四、利用W、SFE-250 bar 及SFE-400 bar 刺激RAW264.7 細胞所得conditioned
medium 對MCF-7、MDA-MB-231 細胞存活之影響。 31

圖目錄
圖一、樟芝水萃取物(W)及超臨界二氧化碳樟芝粉末之水萃取物(SFE-250 bar、
SFE-400 bar) 對乳癌細胞(MCF-7)成長之影響。細胞處理不同濃度的W、
SFE-250 與SFE-400 bar (0 至8 mg/ml) 72 小時後之存活率。 52
圖二、樟芝水萃取物(W)及超臨界二氧化碳樟芝粉末之水萃取物(SFE-250 bar、
SFE-400bar) 對乳癌細胞(MDA-MB-231)成長之影響。細胞處理不同濃度的
W、SFE-250 與SFE-400 bar (0 至8 mg/ml) 72 小時後之存活率。 52
圖三、以樟芝水萃取物(W)及超臨界二氧化碳樟芝粉末之水萃取物(SFE-250 bar、
SFE-400 bar) MCF-7 細胞處理不同濃度的W、SFE-250 與SFE-400 bar (0
至8 mg/ml) 12 小時後細胞凋亡之變化圖。 55
圖四、以樟芝水萃取物(W)及超臨界二氧化碳樟芝粉末之水萃取物(SFE-250 bar、
SFE-400 bar) MDA-MB-231 細胞處理不同濃度的W、SFE-250 與SFE-400
bar (0 至8 mg/ml) 12 小時後細胞凋亡之變化圖。 58
圖五、以W處理MCF-7 細胞以不同濃度0、0.5、1、2、4、8 mg/ml， 12 小時後，
觀察Bcl-2、Bax mRNA 表現量。。。。。 59
圖六、以SFE-250 bar 處理MCF-7 細胞以不同濃度0、0.5、1、2、4、8 mg/ml，
12 小時後，觀察Bcl-2、Bax mRNA 表現量。 60
圖七、以SFE-400 bar 處理MCF-7 細胞以不同濃度0、0.5、1、2、4、8 mg/ml，
12 小時後，觀察Bcl-2、Bax mRNA 表現量。 61
圖八、以W處理MDA-MB-231 細胞以不同濃度0、0.5、1、2、4、8 mg/ml， 12
小時後，觀察Bcl-2、Bax mRNA 表現量。 62
圖九、以SFE-250 bar 處理MDA-MB-231 細胞以不同濃度0、0.5、1、2、4、8 mg/ml，
12 小時後，觀察Bcl-2、Bax mRNA 表現量。 63
圖十、以SFE-400 bar 處理MDA-MB-231 細胞以不同濃度0、0.5、1、2、4、8 mg/ml，
12 小時後，觀察Bcl-2、Bax mRNA 表現量。 64

圖十一、以W之不同濃度( 0、0.5、1、2、4、8 mg/ml ) 處理MCF-7 細胞24 小
時後，觀察Bcl-2、Bax 及pro-caspase-9 蛋白質之表現量。 65
圖十二、以SFE-250 bar 之不同濃度( 0、0.5、1、2、4、8 mg/ml ) 處理MCF-7
細胞24 小時後，觀察Bcl-2、Bax 及pro-caspase-9 蛋白質之表現量。 66
圖十三、以SFE-400 bar 之不同濃度( 0、0.5、1、2、4、8 mg/ml ) 處理MCF-7
細胞24 小時後，觀察Bcl-2、Bax 及pro-caspase-9 蛋白質之表現量。 67
圖十四、以W之不同濃度( 0、0.5、1、2、4、8 mg/ml ) 處理MDA-MB-231 細胞
24 小時後，觀察Bcl-2、Bax 及pro-caspase-9 蛋白質之表現量。 68
圖十五、以SFE-250 bar 之不同濃度( 0、0.5、1、2、4、8 mg/ml ) 處理MDA-MB-231
細胞24 小時後，觀察Bcl-2、Bax 及pro-caspase-9 蛋白質之表現量。69
圖十六、以SFE-400 bar 之不同濃度( 0、0.5、1、2、4、8 mg/ml ) 處理MDA-MB-231
細胞24 小時後，觀察Bcl-2、Bax 及pro-caspase-9 蛋白質之表現量。70
圖十七、以W處理人類癌化纖維母細胞(HT-1080)以不同濃度( 0、0.5、1、2、4、
8 mg/ml ) 24 小時後，觀察MMP-9、MMP-2 蛋白質之表現量。 71
圖十八、以SFE-250 bar 處理人類癌化纖維母細胞(HT-1080)以不同濃度( 0、0.5、
1、2、4、8 mg/ml ) 24 小時後，觀察MMP-9、MMP-2 蛋白質之表現量。
72
圖十九、以SFE-400 bar 處理人類癌化纖維母細胞(HT-1080)以不同濃度( 0、0.5、
1、2、4、8 mg/ml ) 24 小時後，觀察MMP-9、MMP-2 蛋白質之表現量。
73
圖二十、以W處理人類癌化纖維母細胞(HT-1080)以不同濃度( 0、0.5、1、2、4、
8 mg/ml ) 24 小時後，觀察MMP-9、MMP-2 mRNA 之表現量。 74
圖二十一、以SFE-250 bar 處理人類癌化纖維母細胞(HT-1080)以不同濃度( 0、0.5、
1、2、4、8 mg/ml ) 24 小時後，觀察MMP-9、MMP-2 mRNA 之表現
量。 75
圖二十二、以SFE-400 bar 處理人類癌化纖維母細胞(HT-1080)以不同濃度( 0、0.5、
1、2、4、8 mg/ml ) 24 小時後，觀察MMP-9、MMP-2 mRNA 之表現
量。 76
圖二十三、W、SFE-250 bar、SFE-400 bar 水萃物抗氧化能力(重疊圖)；利用流式

細胞儀偵測H2O2 誘導60 分的CHO-K1 細胞，使細胞產生大量ROS 後
給予W、SFE-250 bar、SFE-400 bar 水萃物測試清除自由基ROS 的能
力。濃度為0.5、1、2 mg/ml 。 79
圖二十四、以W不同濃度(0.5、1、2、4、8 mg/ml)處理Raw 264.7 細胞所得上清
液再處理癌細胞MCF-7，72 小時後存活率之影響。 80
圖二十五、以SFE-250 bar 之不同濃度(0.5、1、2、4、8 mg/ml)處理Raw 264.7 細
胞所得上清液再處理癌細胞MCF-7，72 小時後存活率之影響。 80
圖二十六、以SFE-400 bar 之不同濃度(0.5、1、2、4、8 mg/ml)處理Raw 264.7 細
胞所得上清液再處理癌細胞MCF-7，72 小時後存活率之影響。 81
圖二十七、以W之不同濃度(0.5、1、2、4、8 mg/ml)處理Raw 264.7 細胞所得上
清液再處理癌細胞MDA-MB-231，72 小時後存活率之影響。 82
圖二十八、 以SFE-250 bar 之不同濃度(0.5、1、2、4、8 mg/ml)處理Raw 264.7 細
胞所得上清液再處理癌細胞MDA-MB-231，72 小時後存活率之影響。
82
圖二十九、 以SFE-400 bar 之不同濃度(0.5、1、2、4、8 mg/ml)處理Raw 264.7 細
胞所得上清液再處理癌細胞MDA-MB-231，72 小時後存活率之影響。
83

行政院衛生署統計室，2010，99年主要死因分析。
林爽秋，2005，超臨界二氧化碳萃取中草藥及其萃出物對抑制黑色素生成之研
究，國立成功大學化學研究所碩士論文。
吳汶錡，2011，KLF10調控BI-1表現量對高劑量雌激素誘發細胞凋亡之影響，國
立成功大學生命科學所碩士論文。
陳皇琦，2007，樟芝(AC-10)調控乳癌細胞之細胞週期與轉移之機制探討，中國
醫藥大學營養研究所碩士論文。
陳登科，2004，超臨界二氧化碳萃取蛹蟲草蟲草素之探討，朝陽科技大學應用化
學系碩士論文。
郭雅婷，2007，香椿(TSL-1)及樟芝(AC-10)對人類及性骨髓血癌細胞凋亡及細胞
週期的機制探討，中國醫藥大學營養研究所碩士論文。
潘怡瑾，2000，以超臨界二氧化碳萃中藥藥材柏子仁之研究，國立台灣大學化學
工程研究所碩士論文。
Adrover, M.F., Muñoz, M.J., Baez, M.V., Thomas, J., Kornblihtt, A.R., Epstein, A.L.,
Jerusalinsky, D.A. (2010). Characterization of specific cDNA background
synthesis introduced by reverse transcription in RT-PCR assays. Biochimie.
92(12):1839-46.

Ao, Z.H., Xu, Z.H., Lu, Z.M., Xu, H.Y., Zhang, X.M., Dou, W.F.(2009).
Niuchangchih (Antrodia camphorata) and its potential in treating liver diseases.
J. Ethnopharma.121:194-212.
Ashkenazi, A. (2002). Targeting death and decoy receptors of the tumour-necrosis
factor superfamily. Nat Rev Cancer. 2:420-30.
Bauvois, B. (2012) .New facets of matrix metalloproteinases MMP-2 and MMP-9 as
cell surface transducers: Outside-in signaling and relationship to tumor
progression. Biochimica et Biophysica Acta (BBA) - Rev on Cancer. 1825
(1):29-36.
Beatson, C.T. (1896). On treatment of inoperable cases of carcinoma of the mamma:
uggestions for a new method of treatment with illustrative cases. Lancet.
2:104-7.
Bonini, M.G., Rota, C., Tomasi, A., Mason, R.P. (2006) The oxidation of
2',7'-dichlorofluorescin to reactive oxygen species: a self-fulfilling prophesy?
Free Radic Biol Med. 40(6):968-75.
Bratton, S.B., MacFarlane, M., Cain, K., Cohen, G.M. (2000). Protein complexes
activate distinct caspase cascades in death receptor and stress-induced apoptosis.
Ex Cell Res. 256:27-33.
Chen, T., Zheng, W., Wong, S.Y., Yang, F.(2008). Mitochondria-mediated apoptosis
in human breast carcinoma MCF-7 cells induced by a novel selenadiazole
derivative. Biomed & Pharmacotherapy .62 :77-84.

Chen, W., He, F.Y., Li, Y.Q. (2006). The apoptosis effect of hispolon from Phellinus
linteus (Berkeley & Curtis) Teng on human epidermoid KB cells. J
Ethnopharmacol. 105(1-2):280-5.
Chen, Y., Guo, H., Du, Z., Lin, X.Z., Che, Y., Ye, X. (2009). Ecology-based screen
identifies new metabolites from a Cordyceps-colonizing fungus as cancer cell
proliferation inhibitors and apoptosis inducers. Cell Prolif. 42(6):838-47.
Cheng, C.Y., Su, C.C. (2010). Tanshinone IIA inhibits Hep-J5 cells by increasing
calreticulin, caspase-12 and GADD153 protein expression. Int J Mol Med.
26(3):379-85.
Cheng, J.J., Lu, M.K., Lin, C.Y., Chang, C.C. (2011). Characterization and functional
elucidation of a fucosylated 1,6-d-mannogalactan polysaccharide from Antrodia
cinnamomea. Carbohydrate Polymers. 83:545-53.
Cheng, M.H., Cheng, H.T., Lin, S.S., Young, S.C., Pai, C.J., Liao, P.H., Chen, S.C.,
Chou, M.Y., Yang, C.C. (2009). Apoptosis death mode of mitomycin C-treated
HeLa cells and cellular localization of mitomycin C-induced P-glycoprotein.
Drug Chem Toxicol. 32(2):158-68.
Cuzick, J., Forbes, J., Howell, A. (2003). Tamoxifen for breast-cancer prevention.
Lancet.361:177-178.
Du, Y.C., Wu,T.Y., Chang, F.R., Lin, W.Y., Hsu, Y.M., Cheng, F.T., Lue, C.Y., Yen,
M.H., Tsui, Y.T., Chen, H.L., Hou, M.F., Lu, M.C., Wu, Y.C. (2012). Chemical
profiling of the cytotoxic triterpenoid-concentrating fraction and characterization
of ergostane stereo-isomer ingredients from Antrodia camphorate. JPBA. 58:

182-92.
Dumitrescu, R.G .,Cotarla, I. (2005). Understanding breast cancer risk-where do we
stand in. J.Cell Mol Med. 9(1):208-21.
Dumont, P., Ingrassia, L., Rouzeau, S., Ribaucour, F., Thomas, S., Roland, I., Darro,
F., Lefranc, F., Kiss, R. (2007). The Amaryllidaceae isocarbostyril narciclasine
induces apoptosis by activation of the death receptor and/or mitochondrial
pathways in cancer cells but not in normal fibroblasts. Neoplasia. 9(9):766-76.
García-Risco, M.R., Vicente, G., Reglero, G., Fornari, T. (2011). Fractionation of
thyme (Thymus vulgaris L.) by supercritical fluid extraction and
chromatography. J. Supercritical Fluids. 55:949-54.
Ghobrial, I.M., Witzig, T.E., Adjei, A.A. (2005). Targeting apoptosis pathways in
cancer therapy. CA Cancer J. Clin. 55:178-94.
Groenendyk, J., Michalak, M. (2005). Endoplasmic reticulum quality control and
apoptosis. Acta Biochim Pol. 52:381-95.
Huang, G.J., Deng, J.S., Huang, S.S., Shao, Y.Y., Chen, C.C., Kuo, Y.H. (2012).
Protective effect of antrosterol from Antrodia camphorata submerged whole
broth against carbon tetrachloride-induced acute liver injury in mice. Food
Chem .132:709-16.
Hall, J.M., McDonnell, D.P.(1999). The estrogen receptor β-isoform of the human
estrogen receptor modulates Era transcriptional activity and is a key regulator of
the cellular response to estrogens and antiestrogens. Endocrinology.140:5566-78.

Hayashi, A., Silver, S., vna der Westhuizne, N., Donald, J., Parker, C., Fraser, S.
(2003). Treatment of invasive breast carcinoma with ultrasound guided
radiofrequency ablation. Am J Surg. 185(5):429-35.
Hseu, Y.C., Chen, S.C., Liao, J.W., Yang, H.L. (2008) Antrodia camphorate inhibits
proliferation of human breast cancer cells in vitro and in vivo. Food Chem
Toxicol .46: 2680-8.
Hseu, Y.C., Chen, S.C., Tsai, P.C., Chen, C.S., Lu, F.J., Chang, N.W., Yang, H.L.
(2007). Inhibition of cyclooxygenase-2 and induction of apoptosis in
estrogen-nonresponsive breast cancer cells by Antrodia camphorata. Food
Chemi Toxicol. 45: 1107-15.
Hurst, D.R., Welch, D.R.(2011) Metastasis suppressor genes at the interface between
the environment and tumor cell growth. Int Rev Cell Mol Biol. 286:107-80.
Hwang, H.J., Park, H.J., Min, H.Y., Park, E.J., Hong, J.Y., Lee, S.K. (2006).
Inhibitory effects of caffeic acid phenethyl ester on cancer cell metastasis
mediated by the down-regulation of matrix metalloproteinase expression in
human HT-1080 fibrosarcoma cells. J Nutr Biochem. 17(5):356-62.
Isaza-Guzmán, D.M., Carolina, A.O., María, C., Pabón, M., Sergio, I., Arroyave, T.
(2011). Salivary levels of matrix metalloproteinase (MMP) -9 and tissue
inhibitor of matrix metalloproteinase (TIMP)-1: A pilot study about the
relationship with periodontal status and MMP-9 −1562C/T gene promoter
polymorphism. Archives Oral Biology. 56(4):401-11.
Jeong, S.Y., Seol, D.W. (2009). The role of mitochondria in apoptosis. BMB.

41(1):11-22.
Ju, H.K., Lee,H.W., Chung, K.S., Choi, J.H., Cho,J.G., Baek, N.I., Chung, H.G., Lee,
K.T. (2012). Standardized flavonoid-rich fraction of Artemisia princeps
Pampanini cv. Sajabal induces apoptosis via mitochondrial pathway in human
cervical cancer HeLa cells. J. Ethnopharma. 7357:1- 9.
Jung, J.S., Jung, K., Kim, D.H., Kim, H.S. (2012). Selective inhibition of
MMP-9 gene expression by mangiferin in PMA-stimulated human astroglioma
cells: Involvement of PI3K/Akt and MAPK signaling pathways. Pharmac Rese.
66(1):95-103.
Klein, G., Vellenga, E., Fraaije, M.W., Kamps, W.A., de Bont, E.S. (2004). The
possible role of matrix metalloproteinase (MMP-2 and MMP-9) in cancer, e.g.
acute leukemia. Crit Rev Oncol Hematol. 50(2):87-100.
Koshimune, R., Aoe, M., Toyooka, S., Hara, F., Ouchida, M., Tokumo, M., Sano, Y.,
Date, H., Shimizu, N. (2007). Anti-tumor effect of bisphosphonate (YM529) on
non-small cell lung cancer cell lines. BMC Cancer. 12:7-8.
Kumar,B.,Kumar, A.,Ghosh, S., Pandey, B.N., Mishra, K.P., Hazra, B. (2012).
Diospyrin derivative, an anticancer quinonoid, regulates apoptosis at
endoplasmic reticulum as well as mitichondria by modulating cytosolic calcium
in human breast carcinoma cells. Biochem Biophysi Res Commun.417: 903-9.
Larner, S.F., Hayes, R.L., Wang, K.K. (2006). Unfolded protein response after
neurotrauma. J Neurotrauma. 23:807-29.

Liang, C.Z., Zhang, J.K., Shi, Z., Liu, B., Shen, C.Q., Tao, H.M. (2012). Matrine
induces caspase-dependent apoptosis in human osteosarcoma cells in vitro and
in vivo through the upregulation of Bax and Fas/FasL and downregulation of
Bcl-2 . Cancer Chemother Pharmacol. 69:317-31.
Lim, H.K., Choi, Y.A., Park ,W.,Lee, T., Ryu , S.H., Kim, S.Y., Lim, J.R., Kim, J.H.,
Baek ,S.H. (2003). Phosphatidic Acid Regulates Systemic Inflammatory
Responses by Modulating the Akt-Mammalian Target of Rapamycin-p70 S6
Kinase 1 Pathway. J Biol Chem. 278(46):45117-27.
Lindberg, M.K., Moveraer, S., Gao, H., Dahlman-Wright, K., Gustafsson, J.A.,
Ohlsson, C. (2003) Estrogen receptor-β reduces estrogen receptor-α regulated
gene transcripition, supporting a “Ying Yang” relationship between estrogen
receptor α and β in mice. Mol Endocrinol .17(2):203-8.
Liu, C.J., Chiang, C.C., Chiang, B.H. (2012). The elicited two-stage submerged
cultivation of Antrodia cinnamomea for enhancing triterpenoids production and
antitumor activity. J Biochem Engin. 64 :48-54.
Liu, J.J., Huang, T.S., Hsu, M.L., Chen, C.C., Lin, W.S., Lue, F.J., Chang, W.H.
(2004). Antitumor effects of the partially purified polysaccharides from
Antrodia camphorata and the mechanism of its action. Toxi and Applied Pharm.
201:186-93.
Liu, J.J., Lin, M., Yu, J.Y., Liu, B., Bao, J.K. (2010).Targeting apoptotic and
autophagic pathways for cancer therapeutics.Cancer Letters .1-10.
Love, R.R., Duc, N.B., Allred, D.C., Binh, N.C., Dibh, N.V., Kha, N.N., Thuan, T.T,

Thuy, N.V., The, P.N., Cau, T.T., Tung, N.D., Huong, D.T., Quang le, M., Hien,
N.N., Thuong, L., Shen, T.Z., Xin, Y., Zhang, Q., Havighurst, T.C., Yanf, Y.F.,
Hillner, B.E., DeMets, D.L. (2002). Oophorectomy and tamoxifen adjuvant
therapy in premenopausal Vietnamese and Chinese woman with operable breast
cancer. J. Clin Oncol. 20:2559-66.
Luque de Castro, M.D., García-Ayuso, L.E.(1998). Soxhlet extraction of solid
materials: an outdated technique with a promising innovative future. Analytica
Chimica Acta. 369:1-10.
Lynes, E.M., Simmen, T. (2011). Urban planning of the endoplasmic reticulum (ER):
How diverse mechanisms segregate the many functions of the ER. Biochim et
Biophysica Acta. 1813:1893-1905.
McHugh, M.A. and Krukonis, V.J. (1986) Supercritical Fluid Extraction : Principle
and Practice, Butterworth Publishers.
MacPhee, D.J. (2010) Methodological considerations for improving Western blot
analysis. J Pharmacological Toxicol Med. 61:171-7.
Makin, G., Dive, C. (2001). Apoptosis and cancer chemotherapy. Trends Cell Biol.
11:22-6.
Masud, A., Mohapatra, A., Lakhani, S.A., Ferrandino, A., Hakem, R., Flavell, R.A.
(2007). ER stress-induced death of mouse embryonic fibroblasts requires the
intrinsic pathway of apoptosis. J Biol Chem. 19:14132-9.
Nagase, H., Woessner, J.F. (1999). Matrix metalloproteinases. J Biol Chem.

274:21491-4.
Nakagawa, T., Yuan, J. (2000). Cross-talk between two cysteine protease families.
Activation of caspase-12 by calpain apoptosis. J Cell Biol. 150(4):887-94.
Nigam, M., Ranjan, V., Srivastava, S., Sharma, R., Balapure, A.K. (2008).
Centchroman induces G0/G1 arrest and caspase-dependent apoptosis involving
mitochondrial membrane depolarization in MCF-7 and MDA-MB-231 human
breast cancer cells. Life Sci. 82(11-12):577-90.
Papamichail, I., Louli, V., Magoulas, K. (2000). Supercritical fluid extraction of
celery seed oil. J Supercritical fluids. 18(3):231-26.
Parone, P.A., James, D., Martinou, J.C. (2002). Mitochondria: regulating the
inevitable. Biochimie.84(2-3):105-11.
Park, S.E., Yoo, H.S., Jin, C.Y., Hong, S.H., Le, Y.W., Kim, B.W., Lee, S.H., Kim,
W.J., Cho, C.K., Choi, Y.H. (2009). Induction of apoptosis and inhibition of
telomerase activity in human lung carcinoma cells by the water extract of
Cordyceps militaris. Food Chem Toxicol. 47(7):1667-75.
Pathy, N.B., Yip, C.H., Taib, N.A., Hartman, M., Saxena, N., Iau, P., Bulgiba, A.M.,
Lee, S.C., Lim, S.E., Wong, J.E., Verkooijen, H.M. (2011). Breast cancer in a
multi-ethnic Asian setting: results from the Singapore-Malaysia hospital-based
breast cancer registry. Breast. 2:75-80.
Rebbeck,T.R., Kauff, N.D., Domchek, S.M. (2009). Meta-analysis of risk reduction
estimates associated with risk-reducing salpingo-oophorectomy in BRCA1 or

BRCA2 mutation carriers. J Natl Cancer Inst. 101(2):80-7.
Robert, E. (2003). Coleman current and future status of adjuvant therapy for breast
cancer. Cancer. 97:880-886.
Rollin , J., R’egina, S., Vourc’h P., Iochmann, S., Bl’echet, C., Reverdiau, P., Gruel,
Y. (2007). Influence of MMP-2 and MMP-9 promoter polymorphisms on gene
expression and clinical outcome of non-small cell lung cancer. Lung Cancer . 56:
273-80.
Roy, L., LaBoissiere, S., Abdou , E., Thibault, G., Hamel ,N., Taheri ,
M.,Boismenu ,D., Lanoix ,J., Kearney ,R., Paiement, J. (2010). Proteomic
analysis of the transitional endoplasmic reticulum in hepatocellular carcinoma:
An organelle perspective on cancer .Biochim et Biophysica Acta.
1804:1869-1881.
Schiff, P.B., Fant, J., Horwitz, S.B. (1979). Promotion of microtubule assembly in
vitro by Taxol. Nature. 277:665-667.
Searle, J., Kerr, J.F., Bishop, C.J. (1982). Necrosis and apoptosis: distinct modes of
cell death with fundamentally different significance. Pathol Annu. 17:229-59.
Song, F., Zhang, L., Yu, H.X., Lu, R.R., Bao, J.D., Tan, C., Sun, Z. (2012).The
mechanism underlying proliferation-inhibitory and apoptosis-inducing effects of
curcumin on papillary thyroid cancer cells. Food Chem. 132:43-50.
Suboj, P., Babykutty, S., Roshan, D., Gopi, V., Rakesh, S.N., Srinivas, P., Gopala, S.
(2012). Aloe emodin inhibits colon cancer cell migration/angiogenesis by

downregulating MMP-2 & -9, RhoB and VEGF via reduced DNA binding
activity of NF-κB. Europ J Pharm Sci. 45(5):581-91.
Tai, W., Mahato, R., Cheng, C. (2010). The role of HER2 in cancer therapy and
targeted drug delivery. J Cont Rel. 146:264-27.
Tothill, I.E. (2009). Biosensors for cancer markers diagnosis. Semin Cell Dev Biol.
20(1):55-62.
Toyokuni, S., Okamoto, K., Yodoi, J. (1995). Persistent oxidative stress in cancer.
FEBS letts. 358:1-3.
Turner, C., Eskilsson, C.S., Björklund, E. (2002). Collection in analytical-scale
supercritical fluid extraction. J Chrom. 947:1-22.
Wen, C.L., Chang, C.C., Huang, C.S., Kuo, C.L., Hsu, S.L., Deng, J.S., Huang, G.J.
(2011). Anti-inflammatory effects of methanol extract of Antrodia cinnamomea
mycelia both in vitro and in vivo. J Ethnophar. 137:575-84.
Wocssnor, J.E. (1994). The family of matrix metalloproteinases. Ann N Y Acdd Sci.
732:11-30.
Yang, H.L., Kuo, Y.H., Tsai, C.T., Huang, Y.T., Chen, S.C., Chang, H.W., Lin, E.,
Lin, W.H., Hseu, Y.C. (2011). Anti-metastatic activities of Antrodia camphorata
against human breast cancer cells mediated through suppression of the MAPK
signaling pathway. Food Chem Toxicol. 49:290-8.
Yeh, C.T., Rao, Y.K., Yao, C.J.,Yeh, C.F.,Li, C.H., Chuang, S.E.,Luong, H.T.,Lai,
G.M., Tzeng, Y.M. (2009). Cytotoxic triterpenes from Antrodia camphorate

and their mode of action in HT-29 human colon cancer cells. Cancer Letters.
285(1):73-9.
Yolanda, S.V., Cabañas, A., Renuncio, Juan A.R., Pando, C. (2009). Supercritical fluid
extraction of peach (Prunus persica) seed oil using carbon dioxide and ethanol.
J Supercritical fluids. 49(2):167-73.
Zakaria, Y., Rahmat, A., Lope Pihie, A.H.,Abdullah, N.R., Houghton, P.J. (2009).
Eurycomanone induce apoptosis in HepG2 cells via up regulation of p53. Cancer
Cell Int. 9(1):16.
Zhang, L., Yao, H.J., Yu, Y., Zhang, Y., Li, R.J., Ju, R.J., Wang, X.X., Sun, M.G., Shi,
J.F., Lu, W.L. (2012). Mitochondrial targeting liposomes incorporating
daunorubicin and quinacrine for treatment of relapsed breast cancer arising from
cancer stem cells . Biomaterials. 33: 565-82.
Zhao ,D.L., Zou , L.B., Lin , S., Shi ,J.G., Zhu, H.B. (2007). Anti-apoptotic effect of
esculin on dopamine-induced cytotoxicity in the human neuroblastoma
SH-SY5Y cell line. Neuropharma. 53:724-732.
Zhu, L., Wang, H., Wang, L., Wang, Y., Jiang, K., Li, C., Ma, Q., Gao, S., Wang, L.,
Li, W., Cai, M., Wang, H., Niu, G., Lee, S., Yang, W., Fang, X., Chen, X. (2011).
High-affinity peptide against MT1-MMP for in vivo tumor imaging. J Control
Release. 150(3):248-55.
Zong, H.A., Zheng, H.X., Zhen, M.L., Hong, Y.X.,Xiao, M.Z., Wen, F.D. (2009).
Eurycomanone induce apoptosis in HepG2 cells via up regulation of
p53.Cancer Cell Int. 9(1):16.