Cell-type specific and non-redundant anti-proliferative effects of shRNA-mediated Galpha12- and Galpha13 knockdown in lung cancer cell lines
Cell-type specific and non-redundant anti-proliferative effects of shRNA-mediated Galpha12- and Galpha13 knockdown in lung cancer cell lines
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DOI：
10.4236/abb.2014.51011
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发表时间：
2014-01
期刊：
影响因子：
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通讯作者：
Thomas Büch;Marius Grzelinski;Olaf Pinkenburg;T. Gudermann;A. Aigner
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文献类型：
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作者：
Thomas Büch;Marius Grzelinski;Olaf Pinkenburg;T. Gudermann;A. Aigner
在小细胞肺癌细胞中，各种自分泌刺激导致Gq/11和G12/13蛋白的平行激活。Gq/11-PLC-β级联对SCLC细胞中促有丝分裂作用的贡献已得到充分证实，但G12/13信号传导的相关性研究较少。虽然在前列腺癌和乳腺癌中，G12/13激活先前已显示出促进侵袭性而不参与细胞增殖，但我们组的先前数据表明G12/13敲低在小细胞肺癌（SCLC）细胞中具有抗增殖作用。为了进一步研究G12/13依赖性信号在肺肿瘤细胞中的作用，我们在SCLC和NSCLC细胞系中采用了shRNA介导的Gα12、Gα13或两者的靶向作用。慢病毒表达的shRNA导致特异性Gα12和Gα13敲低。值得注意的是，在单一敲除一个家族成员后，未观察到另一个家族成员的反上调。有趣的是，增殖的抑制是细胞系依赖性的。在敲低导致抗增殖的细胞系中，Gα12或Gα13的单敲低足以损害增殖，而Gα12和Gα13的双敲低往往不会进一步增加抗增殖作用。同样，当单次敲低不足以抑制增殖时，在双次敲低中未观察到影响。总之，这些发现表明Gα12和Gα13单独影响细胞增殖，干扰一个家族成员足以产生抗肿瘤作用。
In small cell lung cancer cells, various autocrine stimuli lead to the parallel activation of Gq/11 and G12/13 proteins. The contribution of the Gq/11-PLC-β cascade to the mitogenic effects in SCLC cells is well established, but the relevance of G12/13 signaling is less explored. While in prostate and breast cancer, G12/13 activation has been shown previously to promote invasiveness without being involved in cellular proliferation, previous data from our group indicate anti-proliferative effects of G12/13 knockdown in small cell lung cancer (SCLC) cells. To further investigate the role of G12/13-dependent signaling in lung tumor cells, we employed shRNA-mediated targeting of Gα12, Gα13, or both, in SCLC and NSCLC cell lines. Lentiviral expression of shRNAs resulted in specific Gα12 and Gα13 knockdown. Of note, upon single knockdown of one family member, no counter-upregulation of the other one was observed. Interestingly, inhibition of proliferation was cell line dependent. In cell lines where knock-down led to antiproliferation, single knockdown of either Gα12 or Gα13 was sufficient to impair proliferation and double knockdown of Gα12 and Gα13 tended not to further increase anti-proliferative effects. Likewise, when single knockdown was insufficient for an inhibition of proliferation, no effects were observed in double knockdowns. Taken together, these findings indicate that both Gα12 and Gα13 affect cellular proliferation individually and interference with one family member is sufficient for anti-tumor effects.