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外源性 WW0X 基因表达对原代培养肺癌细胞恶性表型的逆转作用。

Reversing effect of exogenous WWOX gene expression on malignant phenotype of primary cultured lung carcinoma cells.

机构信息

Department of Chest Internal Medicine, Tianjin Chest Hospital, Tianjin 300051, China.

出版信息

Chin Med J (Engl). 2010 Mar 5;123(5):615-20.

Abstract

BACKGROUND

Whether WW domain containing oxidoreductase (WWOX) gene is a tumor-suppressor is still controversial. Some researchers found that the transcription of the WWOX gene was lacking not only in tumor tissues but also in non-tumorous tissues and sometimes in normal tissues. Hence it is important to explore the role of the expression of the exogenous WWOX gene in the proliferation and apoptosis of primary cultured lung carcinoma cells.

METHODS

Lipofection technique was used to determine primary cultured lung carcinoma cells containing the highly expressed exogenous WWOX gene and primary cultured cells with vectors as controls. An animal model of lung cancer was made by subcutaneous implantation of tumor cells into nude mice. RT-PCR, Western blotting, flow cytometry, and TUNEL were used to detect the transcription, expression of the exogenous gene and the effect of the expression of targeted genes on the proliferation and apoptosis of the primary cultured lung carcinoma cells.

RESULTS

The growth, clone formation rate (CFR) ((5.33 +/- 1.53)%) of the primary lung cancer cells transfected with the WWOX gene, tumor size and weight were significantly lower than those of the non-transfected lung cancer cells (CFR: (14.33 +/- 1.53)%) and the primary lung cancer cells transfected with blank plasmids (CFR: (11.00 +/- 1.73)%, P < 0.05). The apoptosis level of primary lung cancer cells transfected with the WWOX gene ((40.72 +/- 5.20)%) was significantly higher than that of the non-transfected lung cancer cells ((2.76 +/- 0.02)%) and the primary lung cancer cells transfected with blank plasmids ((2.72 +/- 0.15)%, P < 0.05).

CONCLUSION

The expression of the exogenous WWOX gene can significantly inhibit the proliferation of lung cancer cells and induce their apoptosis, suggesting that the WWOX gene possesses tumor-suppressing effect.

摘要

背景

WW 结构域包含氧化还原酶(WWOX)基因是否为肿瘤抑制基因仍存在争议。一些研究人员发现,WWOX 基因的转录不仅在肿瘤组织中缺失,而且在非肿瘤组织中有时在正常组织中缺失。因此,探讨外源性 WWOX 基因的表达对原代培养肺癌细胞增殖和凋亡的作用具有重要意义。

方法

采用脂质体转染技术,确定高表达外源性 WWOX 基因的原代培养肺癌细胞和载体对照原代培养细胞。将肿瘤细胞皮下植入裸鼠建立肺癌动物模型。采用 RT-PCR、Western blot、流式细胞术和 TUNEL 检测转录、外源性基因的表达以及目的基因表达对原代培养肺癌细胞增殖和凋亡的影响。

结果

转染 WWOX 基因的原代肺癌细胞生长、克隆形成率(CFR)(5.33±1.53%)、肿瘤大小和重量明显低于未转染肺癌细胞(CFR:14.33±1.53%)和转染空白质粒的原代肺癌细胞(CFR:11.00±1.73%)(P<0.05)。转染 WWOX 基因的原代肺癌细胞凋亡水平(40.72±5.20%)明显高于未转染肺癌细胞(2.76±0.02%)和转染空白质粒的原代肺癌细胞(2.72±0.15%)(P<0.05)。

结论

外源性 WWOX 基因的表达可明显抑制肺癌细胞的增殖并诱导其凋亡,提示 WWOX 基因具有抑瘤作用。

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