[PTX1 in nasopharyngeal carcinoma by RNAi technology].
Wen Zhou, Hong Li, Xiangling Feng, Lei Wang, Bin Zhu, Hui Li, Kai‐Tai Yao, Caiping Ren
Abstract
Wen Zhou, Hong Li, Xiangling Feng, Lei Wang, Bin Zhu, Hui Li, Kai‐Tai Yao, Caiping Ren
Abstract
OBJECTIVE: To explore the expression and the role of PTX1 located at the amplified 12p12-p11 region in nasopharyngeal carcinoma (NPC). METHODS: Semi-quantitative RT-PCR and real-time RT-PCR were applied to detect the expression level of PTX1 in 36 NPC and 8 chronic nasopharyngitis (NP) biopsies. RNAi vector targeting PTX1 was constructed and transfected into NPC cell line 6-10B. The RNAi effect was determined by detecting the expression level of PTX1 in transfected 6-10B cell line. Finally, the cell biological characteristics were compared between transfected 6-10B and parental 6-10B by analyzing the cell cycle distribution and apoptosis status using flow cytometry. RESULTS: RT-PCR and real-time RT-PCR revealed that PTX1 gene was over-expressed in NPC tissues (P<0.05). PTX1 expression was suppressed in NPC cell line 6-10B by approximately 65% by RNAi, confirmed by RT-PCR. The depletion of PTX1 could effectively block the proliferation and induce the apoptosis of NPC cells. CONCLUSION: Blocking the expression of PTX1 on mRNA level changed the characterization of NPC cell line 6-10B by RNAi, suggesting that PTX1 identified in the amplified 12p12-p11 region may be involved in the genesis and development of NPC via promoting the cell proliferation and inhibiting the cell apoptosis.
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OBJECTIVE: To explore the expression and the role of PTX1 located at the amplified 12p12-p11 region in nasopharyngeal carcinoma (NPC). METHODS: Semi-quantitative RT-PCR and real-time RT-PCR were applied to detect the expression level of PTX1 in 36 NPC and 8 chronic nasopharyngitis (NP) biopsies. RNAi vector targeting PTX1 was constructed and transfected into NPC cell line 6-10B. The RNAi effect was determined by detecting the expression level of PTX1 in transfected 6-10B cell line. Finally, the cell biological characteristics were compared between transfected 6-10B and parental 6-10B by analyzing the cell cycle distribution and apoptosis status using flow cytometry. RESULTS: RT-PCR and real-time RT-PCR revealed that PTX1 gene was over-expressed in NPC tissues (P<0.05). PTX1 expression was suppressed in NPC cell line 6-10B by approximately 65% by RNAi, confirmed by RT-PCR. The depletion of PTX1 could effectively block the proliferation and induce the apoptosis of NPC cells. CONCLUSION: Blocking the expression of PTX1 on mRNA level changed the characterization of NPC cell line 6-10B by RNAi, suggesting that PTX1 identified in the amplified 12p12-p11 region may be involved in the genesis and development of NPC via promoting the cell proliferation and inhibiting the cell apoptosis.
Key concepts: Nasopharyngeal carcinoma, RNA interference, Flow cytometry, Transfection, Apoptosis, Molecular biology, Cell culture, Cell growth