Effects of siRNA targeting BMPR-II on the biological activities of human liver cancer cells and its mechanism.
Effects of siRNA targeting BMPR-II on the biological activities of human liver cancer cells and its mechanism.
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靶向BMPR-II的siRNA对人肝癌细胞生物学活性的影响及其机制
DOI：
10.1186/1475-2867-14-55
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发表时间：
2014
影响因子：
5.8
通讯作者：
Wu JB
中科院分区：
文献类型：
--
作者：
Zeng P;Cai S;Zhang JN;Yi FM;Jiang WM;Wu JB
骨形态发生蛋白受体II（BMPR-II）在肿瘤的侵袭和增殖中起重要作用。本研究观察针对骨形态发生蛋白受体II（BMPR-II）的小干扰RNA（siRNA）对人肝细胞生物学活性的影响，并探讨其作用机制。设计并合成了3条针对BMPR-II的siRNA分子序列。本研究分为6组，即I组（正常对照组）、II组（空白对照组）、III组（阴性对照组）和IV-VI组（分别为BMPR-II-siRNA-a、siRNA-b和siRNA-c转染细胞）。测定BMPR-II的mRNA和蛋白水平，以选择BMPR-II沉默的最佳序列。以最佳序列转染肝癌细胞后，检测转染细胞的增殖和侵袭能力，并检测细胞凋亡和细胞周期。分别观察BMPR-II沉默和BMPR-II沉默联合抑制丝裂原活化蛋白激酶（MAPK）信号通路后，VEGF-C蛋白的表达。RT-PCR和Western印迹显示，三种肝癌细胞系中HepG 2中BMPR-II的表达最高（P < 0.01），六组中IV组最低（P < 0.01）。MTT法和transwell法显示，转染后48 h，转染组细胞生长和细胞跨膜数均显著低于对照组（P < 0.05）。流式细胞仪显示，转染后48 h，IV组细胞凋亡率最高，细胞明显阻滞于S期（P < 0.01）。Western blot结果显示，BMPR-Ⅱ沉默后，p-P38（P < 0.01）和VEGF-C（P < 0.01）蛋白表达水平明显降低。PD 98059 + siRNA-BMPR-II-a组和SB 203580 + siRNA-BMPR-II-a组VEGF-C蛋白表达水平明显降低（P < 0.01），SB 203580 + siRNA-BMPR-II-a组VEGF-C蛋白表达水平降低更明显（P < 0.01）。针对BMPR-II的siRNA能显著抑制HepG 2细胞的增殖和侵袭，促进细胞凋亡，阻断HepG 2细胞的S期。其机制可能是BMPR-II沉默通过MAPK/P38和MAPK/ERK 1/2途径，尤其是MAPK/P38下调VEGF-C表达。本研究为肝癌的靶向治疗提供了新的思路。
Bone morphogenetic protein receptor II (BMPR-II) plays an important role in tumor’s invasion and proliferation. In this study, we observed the effects of small interfering RNA (siRNA) targeting bone morphogenetic protein receptor II (BMPR-II) on the biological activities of human liver cells and explore its mechanism. The molecular sequences of three siRNA targeting BMPR-IIwere designed and synthesized. In this study, there were 6 groups including group I (normal control), group II (blank control), group III (negative control) and group IV-VI (BMPR-II-siRNA-a, siRNA-b and siRNA-c-transfected cells, respectively). The levels of mRNA and protein of BMPR-II were determined to select the best sequence for BMPR-II silence. After liver cancer cells were transfected with the best sequence, proliferation and invasion of transfected cells were assessed, and apoptosis and cell cycle were detected. The expressions of mitogen-activated protein kinases (MAPKs) signal pathway-related VEGF-C protein were observed after BMPR-II silence and BMPR-II silence combined with inhibiting MAPKs signal pathway, respectively. RT-PCR and Western blot indicated that BMPR-II expression was the highest in HepG2 among the three liver cancer lines (P < 0.01) and the lowest in group IV among the six groups (P < 0.01). MTT assay and transwell assay revealed that the numbers of cell growth and cell transmembrane were significantly lower in group IV than in control groups 48 h after cells were transfected (P < 0.05). Flow cytometer showed that apoptosis was the highest and cells were significantly blocked in S phase 48 h after cells were transfected in group IV (P < 0.01). Western blot indicated that the protein levels of p-P38 (P < 0.01) and vascular endothelial growth factor-C (VEGF-C) (P < 0.01) were significantly decreased after BMPR-II silence. The protein level of VEGF-C was significantly decreased in PD98059 + siRNA-BMPR-II-a and SB203580 + siRNA-BMPR-II-a groups (P < 0.01), especially in SB203580 + siRNA-BMPR-II-a group (P < 0.01). siRNA targeting BMPR-IIcan markedly inhibit HepG2 proliferation and invasion, promote apoptosis and block HepG2 in S phase. Its mechanism may be that BMPR-II silence down-regulates VEGF-C expression through MAPK/P38 and MAPK/ERK1/2 pathways, especially MAPK/P38. This study provides a new targeted therapy for liver cancer.
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影响因子：
3.2
作者：
O'Sullivan AW;Wang JH;Redmond HP
通讯作者：
Redmond HP
影响因子：
3.3
作者：
de Carvalho, Cyntia H. P.;Nonaka, Cassiano F. W.;Pinto, Leao P.
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影响因子：
2
作者：
Zhang, Wei;Zhang, Mingxin;Zhang, Jin
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影响因子：
6.2
作者：
Guicheux, J;Lemonnier, J;Caverzasio, J
通讯作者：
Caverzasio, J
影响因子：
3.5
作者：
Viñals, F;López-Rovira, T;Ventura, F
通讯作者：
Ventura, F