Wangzaozin A诱导人宫颈癌HeLa细胞胞质骨架重排及细胞增殖抑制
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摘要
该文采用锥虫蓝排染法、吉姆萨染色、流式细胞术及免疫荧光技术研究了wangzaozin A诱导HeLa细胞3种细胞骨架的重排特征以及细胞增殖抑制效应。结果显示,0.8~2.0μmol/L的wangzaozin A可显著抑制HeLa细胞增殖,导致细胞G1期阻滞,使细胞形态变化显著,不对称长条形细胞数量增多,与细胞骨架抑制剂诱导效果相似;wangzaozin A可显著改变细胞微管和角蛋白纤维排布方向,并诱导微管和角蛋白纤维聚合,导致细胞平均荧光强度显著增高,但减少细胞应力纤维数量,使平均荧光强度明显下降,显示了浓度和时间依赖性,该结果与紫杉醇对HeLa细胞的作用效果相似。Western blot显示,wangzaozin A并没有显著改变细胞内角蛋白、β肌动蛋白和α-微管蛋白含量,表明wangzaozin A诱导3种骨架纤维量的改变与其细胞内单体蛋白表达量无关。结果说明,wangzaozin A诱导的细胞骨架重排严重干扰细胞稳态,导致细胞及细胞核形态显著改变、细胞增殖抑制,但该化合物诱导细胞骨架重排的直接靶向位点尚需进一步研究。
The effects of wangzaozin A on cellular proliferation inhibition and the cytoskeleton rearrangement of HeLa cells were investigated by using typan blue exclusion, Giemsa staining, flow cytometry and immunofluorescence assay. The results showed that 0.8-2.0 μmol/L wangzaozin A inhibited the proliferation of HeLa cells, inducing G1 arrest and obvious changes of cellular morphology. The number of cells with asymmetric long strips increased, which was similar to effect of cytoskeleton inhibitors. Wangzaozin A could obviously alter the arrangement direction of microtubule and keratin filaments, and induced the polymerization of them, significantly increased in their mean fluorescence intensity. But it decreased number of stress filaments, and markedly reduced mean fluorescence intensity. These effects were shown concentration-and time-dependent. This phenomenon is similar to the effects of paclitaxel on HeLa cells. Western blot assays suggested that wangzaozin A had on obvious effect on the contents of keratin, β-actin and α-tubulin in HeLa cells, indicating that the changes in the amount of cytokeleton filaments induced by wangzaozin A were not related to the amount of monomeric protein. These results suggested that cytoskeletal rearrangement induced by wangzaozin A severely interfered with cellular homeostasis, leading to significant changes in cell and nuclear morphology and inhibition proliferation of HeLa cells. However, the direct target for cytoskeleton rearrangement induced by this compound still needs further study.
The effects of wangzaozin A on cellular proliferation inhibition and the cytoskeleton rearrangement of HeLa cells were investigated by using typan blue exclusion, Giemsa staining, flow cytometry and immunofluorescence assay. The results showed that 0.8-2.0 μmol/L wangzaozin A inhibited the proliferation of HeLa cells, inducing G1 arrest and obvious changes of cellular morphology. The number of cells with asymmetric long strips increased, which was similar to effect of cytoskeleton inhibitors. Wangzaozin A could obviously alter the arrangement direction of microtubule and keratin filaments, and induced the polymerization of them, significantly increased in their mean fluorescence intensity. But it decreased number of stress filaments, and markedly reduced mean fluorescence intensity. These effects were shown concentration-and time-dependent. This phenomenon is similar to the effects of paclitaxel on HeLa cells. Western blot assays suggested that wangzaozin A had on obvious effect on the contents of keratin, β-actin and α-tubulin in HeLa cells, indicating that the changes in the amount of cytokeleton filaments induced by wangzaozin A were not related to the amount of monomeric protein. These results suggested that cytoskeletal rearrangement induced by wangzaozin A severely interfered with cellular homeostasis, leading to significant changes in cell and nuclear morphology and inhibition proliferation of HeLa cells. However, the direct target for cytoskeleton rearrangement induced by this compound still needs further study.
引文
1 Fletch DA,Mullins RD.Cell mechanics and the cytoskeleton.Nature 2010;463(7280):485-92.
2 Jordan MA,Wilson L.Microtubules and actin filaments:dynamic targets for cancer chemotherapy.Curr Opin Cell Biol 1998;10:123-30.
3 Masoumi S,Harisankar A,Gracias A,Bachinger F,Fufa T,Chandrasekar G,et al.Understanding cytoskeleton regulators in glioblastoma multiforme for therapy design.Drug Des Devel Ther 2016;10:2881-97.
4 陈雪红,苗嫄昕,牛兆山,王春波,耿美玉,韩彦弢.以细胞骨架为靶点的海洋抗肿瘤大环内酯类化合物的研究进展.中国海洋药物杂志(Chen Xuehog,Miao Yuanxin,Niu Zhaoshan,Wang Chunbo,Geng Meiyu,Han Yantao.Research progress of cytoskeleton targeted marine antitumor macrolides.Chinese Journal of Marine Medicine)2010;29(4):59-65.
5 Cho JH,Joo YH,Shin EY,Park EJ,Kim MS.Anticancer effects of colchicine on hypopharyngeal cancer.Anticancer Res 2017;37(11):6269-80.
6 Shah NN,Merchant MS,Cole DE,Jayaprakash N,Bernstein D,Delbrook C,et al.Vincristine sulfate liposomes injection(VSLI,Marqibo?):results from a phase I study in children,adolescents,and young adults with refractory solid tumors or leukemias.Pediatr Blood Cancer 2016;63(6):997-5.
7 Choy H,Akerley W,Devore R.Paclitaxel,carboplatin,and radiation therapy for non-small-cell lung cancer.Lung Cancer1998;12(1):80-6.
8 Singh P,Rathinasamy K,Mohan R,Panda D.Microtubule assembly dynamics:an attractive target for anticancer drugs.IUBMB Life 2008;60(6):368-75.
9 尚海,潘莉,杨澍,陈虹,程卯生.微管蛋白抑制剂的研究进展.药学学报(Shang Hai,Pan Li,Yang Shu,Chen Hong,Chen Maosheng.Progress in the study of tubulin inhibitors.Acta Pharmaceutica Sinica)2010;45(9):1078-88.
10 Peterson JR,Mitchison TJ.Small molecules,big impact:a history of chemical inhibitors and the cytoskeleton.Chem Biol2002;9(12):1275-85.
11 Alushin GM,Lander GC,Kellogg EH,Zhang R,Baker D,Nogales E.High-resolution microtubule structures reveal the structural transitions inαβ-tubulin upon GTP hydrolysis.Cell 2014;157(5):1117-29.
12 Sun KW,Ma YY,Guan TP,Xia YJ,Shao CM,Chen LG,et al.Oridonin induces apoptosis in gastric cancer through Apaf-1,cytochrome c and caspase-3 signaling pathway.World J Gastroenterol 2012;18(48):7144-66.
13 Zhou GB,Kang H,Wang L,Gao L,Liu P,Xie J,et al.Oridonin,a diterpenoid extracted from medicinal herbs,targets AML1-ETO fusion protein and shows potent antitumor activity with low adverse effects on t(8;21)leukemia in vitro and in vivo.Blood2007;109(8):3441-50.
14 Liu CX,Yin QQ,Zhou HC,Wu YL,Pu JX,Xia L,et al.Adenanthin targets peroxiredoxin I and II to induce differentiation of leukemic cells.Nat Chem Biol 2012;8:486-93.
15 Xu HZ,Huang Y,Wu YL,Zhao Y,Xiao WL,Lin QS,et al.Pharicin A,a novel naturalent-kaurene diterpenoid,induces mitotic arrest and mitotoccat,astrophe of cancer cells by interfering with Bub R1function.Cell Cycle 2010;9(14):2897-70.
16 杨相艳,张宜恒,丁兰,汪汉卿.一种天然产物Wangzaozin A的细胞毒活性.物理化学学报(Yang Xiangyan,Zhang Yiheng,Ding Lan,Wang Hanqin.Cytotoxic activity of a natural product wangzaozin A.Acta Phys-Chim Sin)2009;25(9):1749-55.
17 丁兰,张世栋,刘国安,王瀚,杨东娟,王丽.对映–贝壳杉烷二萜化合物Wangzaozin对人胃腺癌细胞SGC-7901生长的影响.西北师范大学学报(自然科学版)(Ding Lan,Zhang Shidong,Liu Guoan,Wang Han,Yang Dongjuan,Wang Li.The effect of Wangzaozin A,an ent-kaurane diterpene on the proliferation of human cancer SGC-7901 cells.Journal of Northwest Normal University,Natural Science)2006;42(3):70-3.
18 丁兰,第五佳丽,田继东,柳志军,杨玲,刘国安,等.对映–贝壳杉烷型二萜化合物Wangzaozin A对人早幼粒白血病HL-60细胞生长抑制和凋亡诱导的作用.第三军医大学学报(Ding Lan,Diwu Jiali,Tian Jidong,Liu Zhijun,Yang Lin,Liu Guoan,Ling Lijun.Effect of an ent-kaurane diterpene Wangzaozin A on proliferation and apoptosis in human promyelocytic leukemia HL-60 cells.Journal of Third Military Medical University)2013;35(8):727-32.
19 丁兰,陈祎平,刘国安,吴炎鹏.Wangzaozin A调节NADPH氧化酶源性活性氧诱导HL-60细胞分化.华中师范大学学报(自然科学版)(Ding Lan,Chen Yiping,Liu Guoan,Wu Yanpeng.(Ding Lan,Chen Yiping,Liu Guoan,Wu Yanpeng.Wangzaozin A regulates NADPH oxidase-derived reactive oxygen species to induce the tifferentiation of HL-60 cells.Zhonghua Normal University Journal,Natural Science)2015;2:252-60.
20 丁兰,陈宗儒,陈祎平,李佩蔚,刘国安.NADPH氧化酶源性活性氧介导epinodosin诱导HL-60细胞分化及细胞骨架重组.中国细胞生物学学报(Ding Lan,Chen Zongru,Chen Yiping,Li Peiwei,Liu Guoan.NADPH Oxidase-derived ROS Mediated the Differentiation and reorganization of cytoskeleton induced by Epinodosin in HL-60 Cells.Chinese Journal of Cell Biology)2016;38(3):273-84.
21 丁兰,李佩蔚,孔花青,陈宗儒,刘国安.对映–贝壳杉烷型二萜Leukamenin E对人早幼粒白血病HL-60细胞分化的诱导作用.中国药房(Ding Lan,Li Peiwei,Kong Huaqing,Chen Zongru,Liu Guoan.Ent-kaurane diterpeneleukamenin Ehuman promyelocytic leukemia HL-60 cellsdifferentiation.China Pharmacy)2016;27(13):1771-4.
22 丁兰,郁开北,刘国安.细胞毒活性二萜Wangzaozin A的晶体结构.高等学校化学学报(Ding Lan,Yu Kaibei,Liu Guoan.Crystal structure for cytotoxic diterpenoids Wangzaozin A.Chemical Journal of Chinese Universities)2005;26(8):1455-8.
23 Andrzej P,Mariusz AS,Anna KW,Lidia G,Agnieszka Z,Alina G.Cytoskeletal reorganization and cell death in mitoxantronetreated lung Cancer cells.Acta Histochem 2016;118(8):784-96.
24 Seung HL,Seung MS,Dong JS,Sun MK,Tack JK,Sukgil S,et al.Epothilones induce human colon cancer SW620 cell apoptosis via the tubulin polymerization-independent activation of the nuclear factor-KB/IKB kinase signal pathway.Mol Cancer Ther2007;6(10):2786-97.
25 Dasgupta A,Sawant MA,Lavhale MS,Krishnapati LS,Ghaskadbi S,Sitasawad SL.AECHL-1,a novel triterpenoid,targets tumor neo-vasculature and impairs the endothelial cell cytoskeleton.Angiogenesis 2015;18(3):283-99.
26 Thomas SL,Zhong D,Zhou W,Malik S,Liotta D,Snyder JP,et al.EF24,a novel curcumin analog,disrupts the microtubule cy-toskeleton and inhibits HIF-1.Cell Cycle 2008;7(15):2409-17.
27 Alberti C.Cytoskeleton structure and dynamic behaviour:quick excursus from basic molecular mechanisms to some implications in cancer chemotherapy.Eur Rev Med Pharmacol Sci 2009;13:13-21.
28 张大永,李子元,施连勇,吴晓明,华维一.ent-贝壳杉烷型二萜化合物的细胞毒活性研究进展.有机化学(Sun Handong,Li Ziyuan,Shi Lianyong,Wu Xiaoming,Hua Weiyi.Advances in research on cytotoxic activity of ent-kaurane diterpenoids.Organic Chemistry)2008;28(11):1911-7.
29 Ding Lan,Zhou Qiyin,Wang Li,Wang Wei,Zhang Shidong,Liu Bo.Comparison of cytotoxicity and DNA damage potential induced by ent-kaurene diterpenoids from Isodon plant.Nat Pro Res 2011;25(15):1402-11.
30 Ding Lan,Hou Qian,Zhou Qiyin,Liu Guoan.Structure-activity relationships of eight ent-kaurene diterpenoids from three Isodon plants.Res Chem Intermediat 2010;36(4):443-52.
31 Knapp LW,O'Guin WM,Sawyer RH.Rearrangement of the keratin cytoskeleton after combined treatment with microtubule and microfilament inhibitors.J Cell Biol 1983;97(6):1788-94.
32 Jose TT,Laura B,Thota G,Kline E,Patel P,Liu Y,et al.Withaferin A inhibits breast cancer invasion and metastasisat subcytotoxic doses by inducing vimentin disassembly and serine 56phosphorylation.Int J Cancer 2011;129:2744-55.
33 Kim SH,Hahm ER,Arlotti JA.Withaferin A inhibits in vivo growth of breast cancer cells accelerated by Notch2 knockdown.Breast Cancer Res Treat 2016;157(1):41-54.
34 王小双,范志宇,王涛,谢亮,余莉,高正祥,等.紫杉醇对大鼠肺血管平滑肌细胞表型和细胞骨架的影响.中华结核和肺癌杂志(Wang Xiaoshuang,Fan Zhiyu,Wang Tao,Xie Liang,Yu Li,Gao Zhengxiang,et al.Effects of paclitaxd on proliferation and transition of phenotype in platelet-derived growth factor-BB induced pulmonary vascular smooth muscle cells.Chin J Tuberc Respir Dis)2012;35(7):511-4.
35 翟中和,王喜忠,丁明孝.细胞生物学,第4版.北京:高等教育出版社(Zhai Zhonghe,Wang Xizhong,Ding Mingxiao.Cell Biology,4th.Beijing:Higher Education Press)2007,198-9.
2 Jordan MA,Wilson L.Microtubules and actin filaments:dynamic targets for cancer chemotherapy.Curr Opin Cell Biol 1998;10:123-30.
3 Masoumi S,Harisankar A,Gracias A,Bachinger F,Fufa T,Chandrasekar G,et al.Understanding cytoskeleton regulators in glioblastoma multiforme for therapy design.Drug Des Devel Ther 2016;10:2881-97.
4 陈雪红,苗嫄昕,牛兆山,王春波,耿美玉,韩彦弢.以细胞骨架为靶点的海洋抗肿瘤大环内酯类化合物的研究进展.中国海洋药物杂志(Chen Xuehog,Miao Yuanxin,Niu Zhaoshan,Wang Chunbo,Geng Meiyu,Han Yantao.Research progress of cytoskeleton targeted marine antitumor macrolides.Chinese Journal of Marine Medicine)2010;29(4):59-65.
5 Cho JH,Joo YH,Shin EY,Park EJ,Kim MS.Anticancer effects of colchicine on hypopharyngeal cancer.Anticancer Res 2017;37(11):6269-80.
6 Shah NN,Merchant MS,Cole DE,Jayaprakash N,Bernstein D,Delbrook C,et al.Vincristine sulfate liposomes injection(VSLI,Marqibo?):results from a phase I study in children,adolescents,and young adults with refractory solid tumors or leukemias.Pediatr Blood Cancer 2016;63(6):997-5.
7 Choy H,Akerley W,Devore R.Paclitaxel,carboplatin,and radiation therapy for non-small-cell lung cancer.Lung Cancer1998;12(1):80-6.
8 Singh P,Rathinasamy K,Mohan R,Panda D.Microtubule assembly dynamics:an attractive target for anticancer drugs.IUBMB Life 2008;60(6):368-75.
9 尚海,潘莉,杨澍,陈虹,程卯生.微管蛋白抑制剂的研究进展.药学学报(Shang Hai,Pan Li,Yang Shu,Chen Hong,Chen Maosheng.Progress in the study of tubulin inhibitors.Acta Pharmaceutica Sinica)2010;45(9):1078-88.
10 Peterson JR,Mitchison TJ.Small molecules,big impact:a history of chemical inhibitors and the cytoskeleton.Chem Biol2002;9(12):1275-85.
11 Alushin GM,Lander GC,Kellogg EH,Zhang R,Baker D,Nogales E.High-resolution microtubule structures reveal the structural transitions inαβ-tubulin upon GTP hydrolysis.Cell 2014;157(5):1117-29.
12 Sun KW,Ma YY,Guan TP,Xia YJ,Shao CM,Chen LG,et al.Oridonin induces apoptosis in gastric cancer through Apaf-1,cytochrome c and caspase-3 signaling pathway.World J Gastroenterol 2012;18(48):7144-66.
13 Zhou GB,Kang H,Wang L,Gao L,Liu P,Xie J,et al.Oridonin,a diterpenoid extracted from medicinal herbs,targets AML1-ETO fusion protein and shows potent antitumor activity with low adverse effects on t(8;21)leukemia in vitro and in vivo.Blood2007;109(8):3441-50.
14 Liu CX,Yin QQ,Zhou HC,Wu YL,Pu JX,Xia L,et al.Adenanthin targets peroxiredoxin I and II to induce differentiation of leukemic cells.Nat Chem Biol 2012;8:486-93.
15 Xu HZ,Huang Y,Wu YL,Zhao Y,Xiao WL,Lin QS,et al.Pharicin A,a novel naturalent-kaurene diterpenoid,induces mitotic arrest and mitotoccat,astrophe of cancer cells by interfering with Bub R1function.Cell Cycle 2010;9(14):2897-70.
16 杨相艳,张宜恒,丁兰,汪汉卿.一种天然产物Wangzaozin A的细胞毒活性.物理化学学报(Yang Xiangyan,Zhang Yiheng,Ding Lan,Wang Hanqin.Cytotoxic activity of a natural product wangzaozin A.Acta Phys-Chim Sin)2009;25(9):1749-55.
17 丁兰,张世栋,刘国安,王瀚,杨东娟,王丽.对映–贝壳杉烷二萜化合物Wangzaozin对人胃腺癌细胞SGC-7901生长的影响.西北师范大学学报(自然科学版)(Ding Lan,Zhang Shidong,Liu Guoan,Wang Han,Yang Dongjuan,Wang Li.The effect of Wangzaozin A,an ent-kaurane diterpene on the proliferation of human cancer SGC-7901 cells.Journal of Northwest Normal University,Natural Science)2006;42(3):70-3.
18 丁兰,第五佳丽,田继东,柳志军,杨玲,刘国安,等.对映–贝壳杉烷型二萜化合物Wangzaozin A对人早幼粒白血病HL-60细胞生长抑制和凋亡诱导的作用.第三军医大学学报(Ding Lan,Diwu Jiali,Tian Jidong,Liu Zhijun,Yang Lin,Liu Guoan,Ling Lijun.Effect of an ent-kaurane diterpene Wangzaozin A on proliferation and apoptosis in human promyelocytic leukemia HL-60 cells.Journal of Third Military Medical University)2013;35(8):727-32.
19 丁兰,陈祎平,刘国安,吴炎鹏.Wangzaozin A调节NADPH氧化酶源性活性氧诱导HL-60细胞分化.华中师范大学学报(自然科学版)(Ding Lan,Chen Yiping,Liu Guoan,Wu Yanpeng.(Ding Lan,Chen Yiping,Liu Guoan,Wu Yanpeng.Wangzaozin A regulates NADPH oxidase-derived reactive oxygen species to induce the tifferentiation of HL-60 cells.Zhonghua Normal University Journal,Natural Science)2015;2:252-60.
20 丁兰,陈宗儒,陈祎平,李佩蔚,刘国安.NADPH氧化酶源性活性氧介导epinodosin诱导HL-60细胞分化及细胞骨架重组.中国细胞生物学学报(Ding Lan,Chen Zongru,Chen Yiping,Li Peiwei,Liu Guoan.NADPH Oxidase-derived ROS Mediated the Differentiation and reorganization of cytoskeleton induced by Epinodosin in HL-60 Cells.Chinese Journal of Cell Biology)2016;38(3):273-84.
21 丁兰,李佩蔚,孔花青,陈宗儒,刘国安.对映–贝壳杉烷型二萜Leukamenin E对人早幼粒白血病HL-60细胞分化的诱导作用.中国药房(Ding Lan,Li Peiwei,Kong Huaqing,Chen Zongru,Liu Guoan.Ent-kaurane diterpeneleukamenin Ehuman promyelocytic leukemia HL-60 cellsdifferentiation.China Pharmacy)2016;27(13):1771-4.
22 丁兰,郁开北,刘国安.细胞毒活性二萜Wangzaozin A的晶体结构.高等学校化学学报(Ding Lan,Yu Kaibei,Liu Guoan.Crystal structure for cytotoxic diterpenoids Wangzaozin A.Chemical Journal of Chinese Universities)2005;26(8):1455-8.
23 Andrzej P,Mariusz AS,Anna KW,Lidia G,Agnieszka Z,Alina G.Cytoskeletal reorganization and cell death in mitoxantronetreated lung Cancer cells.Acta Histochem 2016;118(8):784-96.
24 Seung HL,Seung MS,Dong JS,Sun MK,Tack JK,Sukgil S,et al.Epothilones induce human colon cancer SW620 cell apoptosis via the tubulin polymerization-independent activation of the nuclear factor-KB/IKB kinase signal pathway.Mol Cancer Ther2007;6(10):2786-97.
25 Dasgupta A,Sawant MA,Lavhale MS,Krishnapati LS,Ghaskadbi S,Sitasawad SL.AECHL-1,a novel triterpenoid,targets tumor neo-vasculature and impairs the endothelial cell cytoskeleton.Angiogenesis 2015;18(3):283-99.
26 Thomas SL,Zhong D,Zhou W,Malik S,Liotta D,Snyder JP,et al.EF24,a novel curcumin analog,disrupts the microtubule cy-toskeleton and inhibits HIF-1.Cell Cycle 2008;7(15):2409-17.
27 Alberti C.Cytoskeleton structure and dynamic behaviour:quick excursus from basic molecular mechanisms to some implications in cancer chemotherapy.Eur Rev Med Pharmacol Sci 2009;13:13-21.
28 张大永,李子元,施连勇,吴晓明,华维一.ent-贝壳杉烷型二萜化合物的细胞毒活性研究进展.有机化学(Sun Handong,Li Ziyuan,Shi Lianyong,Wu Xiaoming,Hua Weiyi.Advances in research on cytotoxic activity of ent-kaurane diterpenoids.Organic Chemistry)2008;28(11):1911-7.
29 Ding Lan,Zhou Qiyin,Wang Li,Wang Wei,Zhang Shidong,Liu Bo.Comparison of cytotoxicity and DNA damage potential induced by ent-kaurene diterpenoids from Isodon plant.Nat Pro Res 2011;25(15):1402-11.
30 Ding Lan,Hou Qian,Zhou Qiyin,Liu Guoan.Structure-activity relationships of eight ent-kaurene diterpenoids from three Isodon plants.Res Chem Intermediat 2010;36(4):443-52.
31 Knapp LW,O'Guin WM,Sawyer RH.Rearrangement of the keratin cytoskeleton after combined treatment with microtubule and microfilament inhibitors.J Cell Biol 1983;97(6):1788-94.
32 Jose TT,Laura B,Thota G,Kline E,Patel P,Liu Y,et al.Withaferin A inhibits breast cancer invasion and metastasisat subcytotoxic doses by inducing vimentin disassembly and serine 56phosphorylation.Int J Cancer 2011;129:2744-55.
33 Kim SH,Hahm ER,Arlotti JA.Withaferin A inhibits in vivo growth of breast cancer cells accelerated by Notch2 knockdown.Breast Cancer Res Treat 2016;157(1):41-54.
34 王小双,范志宇,王涛,谢亮,余莉,高正祥,等.紫杉醇对大鼠肺血管平滑肌细胞表型和细胞骨架的影响.中华结核和肺癌杂志(Wang Xiaoshuang,Fan Zhiyu,Wang Tao,Xie Liang,Yu Li,Gao Zhengxiang,et al.Effects of paclitaxd on proliferation and transition of phenotype in platelet-derived growth factor-BB induced pulmonary vascular smooth muscle cells.Chin J Tuberc Respir Dis)2012;35(7):511-4.
35 翟中和,王喜忠,丁明孝.细胞生物学,第4版.北京:高等教育出版社(Zhai Zhonghe,Wang Xizhong,Ding Mingxiao.Cell Biology,4th.Beijing:Higher Education Press)2007,198-9.