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本實驗通過MTT法檢測，得到粉防已堿對HeLa細(xì)胞具有抑制作用，且隨著濃度和時間的增加，抑制率逐漸增高，具有時間、濃度依賴性。為了確定粉防己堿引起的HeLa細(xì)胞增殖抑制作用是否通過凋亡途徑實現(xiàn)，本實驗同時也利用AnnexinV/PI雙染法，經(jīng)流式細(xì)胞儀和激光共聚焦顯微鏡檢測細(xì)胞的凋亡情況，結(jié)果均證實粉防己堿對宮頸癌HeLa細(xì)胞具有促進(jìn)凋亡的作用。細(xì)胞凋亡(apoptosis)，又稱程序性細(xì)胞死亡(programmed cell death, PCD)，是指在一定生理和病理情況下機(jī)體為維持內(nèi)環(huán)境穩(wěn)定,通過基因調(diào)控而使細(xì)胞主動有序的消亡過程,是多細(xì)胞生物的一種生理性細(xì)胞死亡。凋亡不僅對胚胎發(fā)生、器官發(fā)育及保護(hù)機(jī)體穩(wěn)態(tài)等至關(guān)重要，而且在控制細(xì)胞增殖、調(diào)節(jié)腫瘤的發(fā)生和生長中起著重要的作用。凋亡負(fù)調(diào)控腫瘤的發(fā)生發(fā)展，通過凋亡機(jī)體可及時清除過多受損但來不及修復(fù)的或生理上不需要的細(xì)胞。細(xì)胞的凋亡受阻從而導(dǎo)致細(xì)胞群體凋亡和增殖的平衡受到破壞，異常細(xì)胞增殖速度超過凋亡速度，從而形成腫瘤[19]。本實驗結(jié)果證明粉防己堿對宮頸癌HeLa細(xì)胞具有一定的促凋亡作用，有作為宮頸癌藥物治療的可能性。
在今后的實驗中，我們將進(jìn)一步檢測粉防己堿對宮頸癌HeLa細(xì)胞是否具有周期阻滯作用，并檢測與細(xì)胞周期阻滯和凋亡相關(guān)的蛋白，更加深入的了解粉防己堿對HeLa細(xì)胞的增殖抑制的分子機(jī)制。
【參考文獻(xiàn)】
[1]PARKIN DM. Global cancer statistics in the year 2000 [J]. Lancet Oncol, 2001, 2(9):533-543.
[2]PARKIN DM，BRAY F, FERLAY J, et al. Estimating the world cancer burden: Globocan 2000 [J]. Int J Cancer, 2001, 94(2):153-156.
[3]PANG L, HOULT JR. Cytotoxicity to macrophages of tetrandrine, an antisilicosis alkaloid, accompanied by an overproduction of prostaglandins [J]. Biochem Pharmacol, 1997, 53(6):773-782.
[4]ONAI N, TSUNOKAWA T, SUDA M, et al. Inhibitory effects of bisbenzylisoquinoline alkaloids on induction of proinflammatory cytokines, interleukin-1 and tumor necrosis factor-alpha [J]. Planta Med, 1995, 61(6):497-501.
[5]WANG G, LEMOS JR. Tetrandrine:a new ligand to block voltage-dependent Ca2+ and Ca(+)-activated K+ channels [J]. Life Sci, 1995, 56(5):295-306.醫(yī).學(xué).全.在.線m.zxtf.net.cn
[6]CAO ZF. Scavenging effect of tetrandrine of active oxygen radicals [J]. Planta Med, 1996, 62(5):413-414.
[7]韓曉兵，柳友清，盧運萍，等. 粉防已堿對人卵巢癌細(xì)胞株A2780增殖與凋亡的影響 [J].中國腫瘤臨床, 2006, 33(4):190-193.
[8]JIN Q, KANG C, SOH Y, et al. Tetrandrine cytotoxicity and its dual effect on oxidative stress-induced apoptosis through modulating cellular redox states in Neuro 2a mouse neuroblastoma cells [J]. Life Sci, 2002, 71(17):2053-2066.
[9]LAI YL, ChEN YJ, WU TY, et al. Induction of apoptosis in human leukemic U937 cells by tetrandrine [J]. Anticancer Drugs, 1998, 9(1):77-81.
[10]OH SH, LEE BH. Induction of apoptosis in human hepatoblastoma cells by tetrandrine via caspase-dependent Bid cleavage and cytochrome c release [J]. Biochem Pharmacol, 2003, 66(5):725-731.
[11]YOO SM, OH SH, LEE SJ, et al. Inhibition of proliferation and induction of apoptosis by tetrandrine in HepG2 cells [J]. J Ethnopharmacol, 2002, 81(2):225-229.
[12]KUO PL, LIN CC. Tetrandrine-induced cell cycle arrest and apoptosis in Hep G2 cells [J]. Life Sci, 2003, 73(2):243-252.
[13]YASUKAWA K, AKASU M, TAKEUCHI M, et al. Bisbenzylisoquinoline alkaloids inhibit tumor promotion by 12-O-tetradecanoylphorbol-13-acetate in two-stage carcinogenesis in mouse skin [J]. Oncology, 1993, 50(2):137-140.
[14]MENG LH, ZHANG HL, HAYWARD L, et al. Tetrandrine induces early G1 arrest in human colon carcinoma cells by down-regulating the activity and inducing the degradation of G1-S–specific cyclin-dependent kinases and by inducing p53 and p21Cip1 [J]. Cancer Research, 2004, 64:9086-9092.
[15]陳寶安，王為，林國為，等. 漢防己甲素抗腫瘤作用的研究進(jìn)展 [J]. 南京中醫(yī)藥大學(xué)學(xué)報, 2001, 17(2):128-129.
[16]李福全，馬春力，張明遠(yuǎn)，等. 粉防己堿對阿霉素心臟損害大鼠的保護(hù)作 [J]. 佳木斯醫(yī)學(xué)院學(xué)報, 1994, 17(6):12.
[17]CHEN YJ, DAI YS, CHEC BF. The effect of tetrandrine and extracts of centella asiatice on acute radiation dermatitis in rats [J]. Bio Pharm Bull, 1999, 22(7):703.
[18]許文林，錢軍，費霞，等. 漢防己甲素逆轉(zhuǎn)血液系統(tǒng)腫瘤細(xì)胞多藥耐藥的臨床觀察 [J]. 中華血液雜志, 1999, 20(7):383.
[19]周愛枝，胡平，任青，等. 丹參酮2A聯(lián)合順鉑對Hela細(xì)胞生長及p53蛋白表達(dá)水平的影響 [J]. 鄖陽醫(yī)學(xué)院學(xué)報, 2007, 26(6):339-344.
