Regulation of hypoxia-inducible factor-1α on peri-neural invasion in pancreatic cancer through the chemokine receptor CX3CR1
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摘要:
目的 探索低氧诱导因子1α(HIF-1α)和细胞趋化因子受体CX3CR1的相互调节在胰腺癌嗜神经侵袭方面的作用。 方法 通过免疫组织化学实验, 探究在胰腺癌的组织样本中HIF-1α与CX3CR1的相互作用对嗜神经侵袭的作用; 用siRNA双链和过表达质粒沉默/过表达HIF-1α和CX3CR1基因, 然后通过Western blot免疫印迹实验检测HIF-1α、CX3CR1表达的变化。 结果 在胰腺癌组织中, HIF-1α蛋白表达水平与肿瘤直径(F=7.81, P < 0.05), 淋巴结转移(r=0.24, P < 0.01), 肿瘤的病理淋巴结转移阶段(r=0.39, P < 0.05)相关; 在人类胰腺癌细胞系、胰腺癌患者手术标本中, HIF-1α和CX3CR1的表达均具有相关性, 并且与胰腺癌的嗜神经侵袭相关联。 结论 HIF-1α和CX3CR1的相互作用可以调节胰腺癌的嗜神经侵袭。 Abstract:Objective This study aimed to elucidate the mechanism underlying the reciprocal regulation between hypoxia-inducing factor-1α(HIF-1α) and the chemokine receptor CX3CR1 on the neural invasion of pancreatic cancer cells. Methods Immunohistochemistry was used to examine the interaction between HIF-1α and CX3CR1 in tissue specimens of pancreatic cancer, and the effect of this interaction on peri-neural invasion(PNI).Then, HIF-1α and CX3CR1 gene were knocked down or overexpressed by the construction of siRNA duplexes/ plasmids to estimate their correlation.The reciprocal regulation of HIF-1α and CX3CR1 was assessed by Western blot assay. Results HIF-1 α was involved in PNI in pancreatic cancer specimens.HIF-1α expression was correlated with clinicopathological parameters in pancreatic cancer patients.HIF-1α and CX3CR1 were correlatively expressed in human pancreatic cancer cell lines and tissue specimens, and both were associated with PNI in pancreatic cancer. Conclusion The reciprocal regulation of HIF-1α and CX3CR1 modulates the neurotropism of pancreatic cancer. -
Key words:
- pancreatic cancer /
- perineural invasion /
- hypoxia inducible factor-1(α subunit) /
- CX3CR1
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图 1 胰腺癌手术标本中HIF-1α和CX3CR1的表达参与了胰腺癌的神经周围浸润
Figure 1. Involvement of HIF-1αand CX3CR1 expression in the peri-neural invasion of pancreatic cancer
图 2 胆固醇包裹的siHIF-1α处理对注射胰腺癌细胞Mia-PaCa-2成瘤裸鼠的影响
Figure 2. Immunohistochemistry of tumors from Mia-PaCa-2 cells trans-planted into nude mice to test the effects of drug delivery of cholester-ol-conjugated siControl(left)or siHIF1a#2(magnification×400)(right)
图 3 胆固醇包裹的siHIF-1α处理对注射胰腺癌细胞Mia-PaCa-2成瘤裸鼠的影响
Figure 3. Immunohistochemistry of tumors originated from MiaPaCa2 cells that were transplanted in nude mice to test the effects of delivery ofcholesterol-conjugated siControl(left)or siHIF1a#2(magnification: ×400)(right)
图 4 HIF-1α和CX3CR1在几种胰腺癌细胞系中的基础表达
Figure 4. Expression of OB-R and HIF-1αin pancreatic cancer cells as-sessed by Western blot
图 5 用HIF-1αsiRNA(50 nM)和pEGFP-C1-CX3CR1过表达质粒(1μg)共转染Mia-PaCa-2细胞, 48 h后, 用CX3CL1(200 ng/mL)处理细胞20 min, 然后用Western blot印迹法来评估蛋白的表达
Figure 5. Pancreatic cancer cells Mia-PaCa-2 were co-transfected with HIF-1α#2 siRNA(50 nM)and pEGFP-C1-CX3CR1 overexpression plasmids(1μg).After 48 h, cells were treated with CX3CL1(200 ng/mL)for 20 min, and the protein expression levels were assessed by West-ern blot
表 1 在胰腺癌中HIF-1α蛋白表达水平与临床病理特征的相关性
Table 1. Correlations between clinicopathological features and HIF-1α expression in pancreatic cancer
表 2 在胰腺癌中HIF-1α和CX3CR1之间的相关性
Table 2. Correlations between HIF-1α expression and CX3CR1 in pancreatic cancer
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