本实验通过MTT法检测,得到粉防已碱对HeLa细胞具有抑制作用,且随着浓度和时间的增加,抑制率逐渐增高,具有时间、浓度依赖性。为了确定粉防己碱引起的HeLa细胞增殖抑制作用是否通过凋亡途径实现,本实验同时也利用AnnexinV/PI双染法,经流式细胞仪和激光共聚焦显微镜检测细胞的凋亡情况,结果均证实粉防己碱对宫颈癌HeLa细胞具有促进凋亡的作用。细胞凋亡(apoptosis),又称程序性细胞死亡(programmed cell death, PCD),是指在一定生理和病理情况下机体为维持内环境稳定,通过基因调控而使细胞主动有序的消亡过程,是多细胞生物的一种生理性细胞死亡。凋亡不仅对胚胎发生、器官发育及保护机体稳态等至关重要,而且在控制细胞增殖、调节肿瘤的发生和生长中起着重要的作用。凋亡负调控肿瘤的发生发展,通过凋亡机体可及时清除过多受损但来不及修复的或生理上不需要的细胞。细胞的凋亡受阻从而导致细胞群体凋亡和增殖的平衡受到破坏,异常细胞增殖速度超过凋亡速度,从而形成肿瘤[19]。本实验结果证明粉防己碱对宫颈癌HeLa细胞具有一定的促凋亡作用,有作为宫颈癌药物治疗的可能性。
在今后的实验中,我们将进一步检测粉防己碱对宫颈癌HeLa细胞是否具有周期阻滞作用,并检测与细胞周期阻滞和凋亡相关的蛋白,更加深入的了解粉防己碱对HeLa细胞的增殖抑制的分子机制。
【参考文献】
[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.医.学.全.在.线www.lindalemus.com
[6]CAO ZF. Scavenging effect of tetrandrine of active oxygen radicals [J]. Planta Med, 1996, 62(5):413-414.
[7]韩晓兵,柳友清,卢运萍,等. 粉防已碱对人卵巢癌细胞株A2780增殖与凋亡的影响 [J].中国肿瘤临床, 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]陈宝安,王为,林国为,等. 汉防己甲素抗肿瘤作用的研究进展 [J]. 南京中医药大学学报, 2001, 17(2):128-129.
[16]李福全,马春力,张明远,等. 粉防己碱对阿霉素心脏损害大鼠的保护作 [J]. 佳木斯医学院学报, 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]许文林,钱军,费霞,等. 汉防己甲素逆转血液系统肿瘤细胞多药耐药的临床观察 [J]. 中华血液杂志, 1999, 20(7):383.
[19]周爱枝,胡平,任青,等. 丹参酮2A联合顺铂对Hela细胞生长及p53蛋白表达水平的影响 [J]. 郧阳医学院学报, 2007, 26(6):339-344.