γ-突触核蛋白在结直肠癌组织中的表达及其对结直肠癌细胞系 HCT116 的作用。
Expression of gamma-synuclein in colorectal cancer tissues and its role on colorectal cancer cell line HCT116.
机构信息
Shanghai Institute of Digestive Surgery, No. 197 Ruijin Second Road, Shanghai 200025, China.
出版信息
World J Gastroenterol. 2009 Oct 28;15(40):5035-43. doi: 10.3748/wjg.15.5035.
AIM
To investigate the expression pattern of gamma-synuclein in colorectal cancer (CRC) tissues, and to study the effects of gamma-synuclein on CRC cell line HCT116 biological features in vitro.
METHODS
The expression pattern of gamma-synuclein was determined in 54 CRC tissues and 30 tumor-matched nonneoplastic adjacent tissues (NNAT) 5 cm away from the tumor via real-time quantitative reverse transcription PCR (RT-PCR) and immunohistochemistry. The relationship between gamma-synuclein protein expression and clinicopathological factors of CRC tissues was analyzed. Three small interfering RNA (siRNA) targeting gamma-synuclein mRNA plasmids were constructed and transfected into the CRC cell line HCT116. The stable cell lines were selected with G-418 for 28 d, and the biological features of these cells were examined by cell growth curve, soft agar assay, and cell migration and invasion assays in vitro.
RESULTS
The expression of gamma-synuclein mRNA and protein was much higher in CRC tissue samples than in NNAT samples (P = 0.02, P = 0.036). There was a significant correlation between the gamma-synuclein protein expression and clinical stage and lymph node involvement of CRC (P = 0.02, P = 0.033). In functional analysis we found that down-regulation of gamma-synuclein expression in HCT116 cells could inhibit the growth, colony formation rate, and migration and invasion ability of HCT116 cells.
CONCLUSION
Increased expression of gamma-synuclein in CRC tissues and the biological effects of reduced gamma-synuclein expression on HCT116 cells suggest that gamma-synuclein may play a positive role in the progression of CRC.
目的
研究γ-突触核蛋白在结直肠癌（CRC）组织中的表达模式，并研究γ-突触核蛋白在体外对 CRC 细胞系 HCT116 生物学特性的影响。
方法
通过实时定量逆转录 PCR（RT-PCR）和免疫组织化学法检测 54 例 CRC 组织和 30 例距肿瘤 5cm 的肿瘤匹配非肿瘤相邻组织（NNAT）中γ-突触核蛋白的表达模式。分析γ-突触核蛋白蛋白表达与 CRC 组织临床病理因素的关系。构建了 3 个靶向 γ-突触核蛋白 mRNA 的小干扰 RNA（siRNA）质粒，并转染 CRC 细胞系 HCT116。用 G-418 筛选稳定转染细胞系 28d，通过细胞生长曲线、软琼脂试验和细胞迁移及侵袭试验体外检测这些细胞的生物学特性。
结果
CRC 组织样本中γ-突触核蛋白 mRNA 和蛋白的表达均明显高于 NNAT 样本（P = 0.02，P = 0.036）。γ-突触核蛋白蛋白表达与 CRC 的临床分期和淋巴结受累显著相关（P = 0.02，P = 0.033）。在功能分析中发现，下调 HCT116 细胞中的γ-突触核蛋白表达可抑制 HCT116 细胞的生长、集落形成率以及迁移和侵袭能力。
结论
CRC 组织中γ-突触核蛋白表达增加，以及降低γ-突触核蛋白表达对 HCT116 细胞的生物学作用提示，γ-突触核蛋白可能在 CRC 的进展中发挥积极作用。
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