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中华肝脏外科手术学电子杂志 ›› 2014, Vol. 03 ›› Issue (06) : 380 -383. doi: 10.3877/cma.j.issn.2095-3232.2014.06.012

所属专题: 文献

基础研究

siRNA沉默蛋白激酶Cε基因抑制胆管细胞癌生长及其作用机制
郭明明1, 刘江辉1, 蔡锐彬1, 王晶2,(), 陈斌3,()   
  1. 1. 510080 广州,中山大学附属第一医院急诊科
    2. 510080 广州,中山大学附属第一医院妇产科
    3. 510080 广州,中山大学附属第一医院肝胆外科
  • 收稿日期:2014-10-02 出版日期:2014-12-10
  • 通信作者: 王晶, 陈斌
  • 基金资助:
    国家自然科学基金(81302550)

Protein kinase C-epsilon gene silenced by siRNA inhibits the development of cholangiocellular carcinoma and its mechanism

Mingming Guo1, Jianghui Liu1, Ruibin Cai1, Jing Wang2,(), Bin Chen3,()   

  1. 1. Department of Emergency, the First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510080, China
引用本文:

郭明明, 刘江辉, 蔡锐彬, 王晶, 陈斌. siRNA沉默蛋白激酶Cε基因抑制胆管细胞癌生长及其作用机制[J]. 中华肝脏外科手术学电子杂志, 2014, 03(06): 380-383.

Mingming Guo, Jianghui Liu, Ruibin Cai, Jing Wang, Bin Chen. Protein kinase C-epsilon gene silenced by siRNA inhibits the development of cholangiocellular carcinoma and its mechanism[J]. Chinese Journal of Hepatic Surgery(Electronic Edition), 2014, 03(06): 380-383.

目的

探讨小干扰RNA(siRNA)沉默蛋白激酶Cε (PKCε)基因抑制胆管细胞癌的生长及其作用机制。

方法

分别将PKCε-siRNA及阴性对照(NC)-siRNA转染至人胆管细胞癌QBC939细胞中,设立PKC组和NC组。另设未转染对照组(CTRL组)。采用细胞计数试剂盒(CCK)-8法检测细胞增殖抑制率,流式细胞术检测细胞凋亡率,蛋白质印迹法检测PKCε、survivin蛋白表达情况。3组比较采用单因素方差分析,两两比较采用LSD-t检验。

结果

PKC组细胞24、48、72 h的细胞增殖抑制率逐渐增高,分别为(7.52±0.33)%、(15.28±0.20)%和(37.12±0.45)%,与CTRL组比较差异有统计学意义(LSD-t =15.37,27.12,35.05;P<0.05)。PKC组细胞凋亡率(56.9±6.1)%明显高于CTRL组的(12.5±1.3)% (LSD-t=28.55,P<0.05)。PKC组PKCε、survivin蛋白表达水平较NC组、CTRL组降低。

结论

siRNA沉默PKCε基因可能通过下调survivin蛋白表达,促进细胞凋亡从而抑制胆管细胞癌的生长。

Objective

To investigate the role of protein kinase C-epsilon (PKCε) gene silenced by small interfering ribonucleic acid (siRNA) in inhibiting the development of cholangiocellular carcinoma and its mechanism.

Methods

Human cholangiocellular carcinoma QBC939 cells were respectively transfected using PKCε-siRNA and negative control (NC)-siRNA to establish PKC group and NC group. And untransfected control (CTRL) group was established. Cell counting kit (CCK)-8 assay was used to define the cell proliferation inhibition rate. Cell apoptosis rate was detected by flow cytometery. The expressions of protein PKCε and survivin were detected by Western blot. The comparison of three groups was conducted using one way analysis of variance and pairwise comparison using LSD-t test.

Results

The cell proliferation inhibition rates at 24, 48, 72 h [(7.52±0.33)%, (15.28±0.20)%, (37.12±0.45)% ] increased gradually in PKC group, where significant difference was observed compared with those in CTRL group (LSD-t=15.37, 27.12, 35.05; P<0.05). The cell apoptosis rate was (56.9±6.1)% in PKC group, which was significantly higher compared with that in CTRL group [(12.5±1.3) % ] (LSD-t=28.55, P<0.05). The expressions of protein PKCε and survivin decreased in PKC group compared with those in NC group and CTRL group.

Conclusion

PKCε gene silenced by siRNA may inhibit the development of cholangiocellular carcinoma through down-regulating the expression of protein survivin and promoting cell apoptosis.

图1 Western blot法检测QBC939人胆管细胞癌细胞PKCε、survivin蛋白表达
[1]
Nomura R, Fujii H, Abe M, et al. Mesothelin expression is a prognostic factor in cholangiocellular carcinoma[J]. Int Surg, 2013, 98(2):164-169.
[2]
Hu C, Cheng C, Pan S, et al. PKC mediates fluctuant ERK-paxillin signaling for hepatocyte growth factor-induced migration of hepatoma cell HepG2[J]. Cell Signal, 2013, 25(6):1457-1467.
[3]
Jain K, Basu A. The multifunctional protein kinase C-ε in cancer development and progression[J]. Cancers, 2014, 6(2):860-878.
[4]
Körner C, Keklikoglou I, Bender C, et al. MicroRNA-31 sensitizes human breast cells to apoptosis by direct targeting of protein kinase C epsilon (PKC epsilon)[J]. J Biol Chem, 2013, 288(12):8750-8761.
[5]
Guo Y, Liang X, Lu M, et al. Mammalian target of rapamycin as a novel target in the treatment of hepatocellular carcinoma[J]. Hepatogastroenterology, 2009, 57(101):913-918.
[6]
Ye X, Guo Y, Zhang Q, et al. βKlotho suppresses tumor growth in hepatocellular carcinoma by regulating Akt/GSK-3β/Cyclin D1 signaling pathway[J]. PloS One, 2013, 8(1):e55615.
[7]
Guo Y, Chen W, Wang W, et al. Simultaneous diagnosis and gene therapy of immuno-rejection in rat allogeneic heart transplantation model using a T-cell-targeted theranostic nanosystem[J]. ACS Nano, 2012, 6(12):10646-10657.
[8]
Wang J, Ou J, Guo Y, et al. TBLR1 is a novel prognostic marker and promotes epithelial-mesenchymal transition in cervical cancer[J]. Br J Cancer, 2014, 111(1):112-124.
[9]
Wang C, Guo Y, Wang J, et al. The suppressive role of SOX7 in hepatocarcinogenesis[J]. PloS One, 2014, 9(5):e97433.
[10]
Diaz-Cueto L, Arechavaleta-Velasco F, Diaz-Arizaga A, et al. PKC signaling is involved in the regulation of progranulin (acrogranin/PC-cell-derived growth factor/granulin-epithelin precursor) protein expression in human ovarian cancer cell lines[J]. Int J Gynecol Cancer, 2012, 22(6):945-950.
[11]
Dudek H, Wong DH, Arvan R, et al. Knockdown of β-catenin with dicer-substrate siRNAs reduces liver tumor burden in vivo[J]. Mol Ther, 2014, 22(1):92-101.
[12]
法镇中,江勇,谭玉林,等.慢病毒介导siRNA沉默EGFR基因及其对胰腺癌细胞增殖和凋亡的影响[J/CD].中华肝脏外科手术学电子杂志,2013, 2(2):113-118.
[13]
Huang B, Cao K, Li X, et al. The expression and role of protein kinase C (PKC) epsilon in clear cell renal cell carcinoma[J]. J Exp Clin Cancer Res, 2011, 30(1):88.
[14]
Gobbi G, Masselli E, Micheloni C, et al. Hypoxia-induced down-modulation of PKCepsilon promotes trail-mediated apoptosis of tumor cells[J]. Int J Oncol, 2010, 37(3):719-729.
[15]
Chen J, Giridhar KV, Zhang L, et al. A protein kinase C/protein kinase D pathway protects LNCaP prostate cancer cells from phorbol ester-induced apoptosis by promoting ERK1/2 and NF-{kappa}B activities[J]. Carcinogenesis, 2011, 32(8):1198-1206.
[16]
Aziz MH, Shen H, Maki CG. Glucocorticoid receptor activation inhibits p53-induced apoptosis of MCF10Amyc cells via induction of protein kinase Cepsilon[J]. J Biol Chem, 2012, 287(35):29825-29836.
[17]
Abd EI-Hakim TF, El-Shafie MK, Abdou AG, et al. Value of urinary survivin as a diagnostic marker in bladder cancer[J]. Anal Quant Cytopathol Histpathol, 2014, 36(3):121-127.
[18]
Salzano G, Riehle R, Navarro G, et al. Polymeric micelles containing reversibly phospholipid-modified anti-survivin siRNA: a promising strategy to overcome drug resistance in cancer[J]. Cancer Lett, 2014, 343(2):224-231.
[19]
Wenying Z, Zhaoning J, Zhimin Y, et al. Survivin siRNA inhibits gastric cancer in nude mice[J]. Cell Biochem Biophys, 2012, 62(2):337-341.
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