Mechanism of 9-cisRA in Inhibiting the Proliferation of Thyroid Squamous Cell Carcinoma Cell SW579
-
摘要:
目的 观察9-顺式维甲酸(9-cis-retinoic acid, 9-cisRA)对甲状腺鳞癌细胞株SW579细胞RARβ、p21、CDK4、CyclinDl表达的影响, 进而探讨9-cisRA的抗癌作用机制。 方法 分别以终浓度为1×10-7、1×10-6、5×10-6、1×10-5mol/L 9-cisRA作用SW579细胞, 各组细胞均在培养24 h后加药, 继续培养48 h。用RT-PCR方法检测SW579细胞RARβ、p21、CDK4、CyclinD1的mRNA表达; 用Western Blot检测SW579细胞RARβ、p21、CDK4、CvclinD1的蛋白表达。 结果 9-cisRA作用后, RT-PCR检测结果表明, 经不同浓度9-cisRA处理的细胞RARB、p21的mRNA表达水平均显著上调; CDK4的mRNA表达水平无明显变化; CyclinD1的表达水平明显下调; Western Blot检测结果表明经不同浓度9-cisRA处理的细胞RARβ、p21蛋白表达水平均显著上调; CDK4蛋白表达水平无明显变化; CyclinD1蛋白表达水平明显下调。 结论 9-cisRA在一定浓度范围内可能通过上调RARβ、p21的表达进而下调细胞周期蛋白CvclinD1的表达, 从而抑制细胞增殖。 Abstract:Objectives The current study aims to observe the effect of 9-cisRA in the expressions of RARP, p21, CDK4, and CyinDl genes in the thyroid squamous cell carcinoma cells(SW579) to explore its mechanism. Methods The SW579 cells were treated with different doses of 9-cisRA(final concentration of 1 x 10-7 mol/L, 1×10-6 mol/L, 5x10-6 mol/L, and 1×10-5 mol/L).The drug was added in the cells of all groups 24 h after the culture.The cells were then cultured for another 48 h.The RARβ, p21, CDK4, and CyclinDl mRNA expression levels were determined via RT-PCR, whereas the expression of RARβ, p21, CDK4, and CyclinDl protein were determined using Western blot. Results The mRNA expression of RARβand p21 increased dramatically after the cells were treated with 9-cisRA, whereas the expression of CDK4 did not increase.In addition, the mRNA expression of CyclinDl dramatically decreased, whereas the protein expression of RARβand p21 significantly increased.No distinct change in the protein expression of CDK4 was observed.The CyclinDl protein expression was dramatically reduced. Conclusion The drug 9-cisRA can inhibit the proliferation of thyroid squamous cell carcinoma cells in a dose-dependent manner.The mechanism of the effect may be related to the increase of RARβand p21mRNA and protein expression, as well as the decrease of CyclinDl mRNA and protein expression. -
图 1 各组细胞CDK4 mRNA表达水平
M：Marker；1：空白对照组；2：乙醇对照组；3：9-cisRA 1×10-7mol/L组；4：9-cisRA 1×10-6mol/L组；5：9-cisRA 5×10-6mol/L组；6：9-cisRA 1×10-5mol/L组
Figure 1. mRNA expression of CDK4 in different groups
图 2 各组细胞CyclinD1 mRNA表达水平
M：Marker；1：空白对照组；2：乙醇对照组；3：9-cisRA 1×10-7 mol/L组；4：9-cisRA 1×10-6mol/L组；5：9-cisRA 5×10-6mol/L组；6：9-cisRA 1×10-5mol/L组
Figure 2. mRNA expression of CyclinD1 in different groups
图 3 各组细胞RARβ mRNA表达水平
M：Marker；1：空白对照组；2：乙醇对照组；3：9-cisRA 1×10-7mol/L组；4：9-cisRA 1×10-6mol/L组；5：9-cisRA 5×10-6mol/L组；6：9-cisRA 1×10-5mol/L组
Figure 3. mRNA expression of RARβ in different groups
图 4 各组细胞p21 mRNA表达水平
M：Marker；1：空白对照组；2：乙醇对照组；3：9-cisRA 1×10-7mol/L组；4：9-cisRA 1×10-6mol/L组；5：9-cisRA 5×10-6mol/L组；6：9-cisRA 1×10-5mol/L组
Figure 4. mRNA expression of p21 in different groups
图 5 CDK4、CyclinD1、RARβ、p21蛋白表达
1：对照组；2：9-cisRA 1×10-7组；3：9-cisRA 1×10-6组；4：9-cisRA 5×10-6组；5：9-cisRA 1×10-5组
Figure 5. Protein expression of CDK4, CyclinD1, RARβ, and p21 in different groups
表 1 CDK4、CyclinD1、RARβ、p21 mRNA的表达水平 x±s，n=3
Table 1. mRNA expression of CDK4, CyclinD1, RARß and p21(x±s, n=3)
表 2 CDK4、CyclinD1、RARβ、p21蛋白的表达水平 x±s，n=3
Table 2. Protein expression of CDK4, CyclinD1, RARβ, and p21(x±s, n=3)
-
[1] Schmutzler C, Hoang—Vu C, R(u|¨)ger B, et al. Human thyroid carcinoma cell lines show different retinoic add receptor repertoires and retinoid responses[J]. Eur J Endocrinol, 2004, 150(4): 547-556. http://citeseerx.ist.psu.edu/viewdoc/download;jsessionid=1DD38ACBC092871BD943874243C38618?doi=10.1.1.323.3448&rep=rep1&type=pdf [2] 范会瑜, 肖建英, 李男, 等. 维甲酸抑制甲状腺鳞癌细胞株生长增殖作用机制的探讨[J]. 中国地方病防治杂志, 2009, 24(2): 91-95. https://www.cnki.com.cn/Article/CJFDTOTAL-DYBF200902004.htm [3] 付锦艳, 潘晓琳, 杨安强, 等. 细胞周期调控相关蛋白CyclinD1、GDK4和pRb在新疆维吾尔族妇女宫颈癌中的表达及意义[J]. 肿瘤防治研究, 2009, 36(11): 969-972. doi: 10.3971/j.issn.1000-8578.2009.11.18 [4] 周艳红, 徐瑞成, 呼文亮, 等. 9-顺式维甲酸对胃癌细胞周期及周期因子表达的影响[J]. 肿瘤, 2007, 27(2): 92-95. https://www.cnki.com.cn/Article/CJFDTOTAL-ZZLL200702003.htm [5] 王卫理, 周士福, 费伯健, 等. 乳腺癌维甲酸受体表达的临床研究[J]. 南京医科大学学报, 2005, 25(2): 118-120. https://www.cnki.com.cn/Article/CJFDTOTAL-NJYK200502013.htm [6] 徐瑞成周艳红, 呼文亮. 9-顺式维甲酸诱导胃癌MGC803细胞周期阻滞和凋亡的作用机制[J]. 癌症, 2006, 25(12): 1483-1487. https://www.cnki.com.cn/Article/CJFDTOTAL-AIZH200612006.htm [7] Choi EJ, Whang YM, Kim SJ, et al. Combinational treatment with retinoic add derivatives in non—small cell lung carcinoma in vitro[J]. J Korean Med Sd, 2007, 22(Suppl): S52-S60. doi: 10.3346/jkms.2007.22.S.S52 [8] Mernitz H, Smith DE, Zhu AX, et al. 9—ds—Retinoic add inhibition of lung carcinogenesis in the A/J mouse model is accompanied by increased expression of RAR—beta but no change in cydooxygenase—2[J]. Cancer Lett, 2006, 244(1): 101-108. doi: 10.1016/j.canlet.2005.12.001 [9] Bonofiqho D, Cione E, Qi H, et al. Combined low doses of PPARgamma and RXR ligands trigger an intrinsic apoptotic pathway in human breast cancer cells[J]. Am J Pathol, 2009, 175(3): 1270-1280. doi: 10.2353/ajpath.2009.081078 [10] Tanaka T, Suh KS, Lo AM, et al. p21WAF1/CIPl is a common transcriptional target of retinoid receptors: pleiotropic regulatory mechanism through retinoic add receptor(RAR)/retinoid X receptor (RXR) heterodimer and RXR/RXR homodimer[J]. J Biol Chem, 2007, 282(41): 29987-29997. doi: 10.1074/jbc.M701700200 -
下载:
点击查看大图
计量
- 文章访问数: 35
- HTML全文浏览量: 24
- PDF下载量: 0
- 被引次数: 0
