Application of the dual-micropipet technique to the measurement of tumor cell locomotion.
Application of the dual-micropipet technique to the measurement of tumor cell locomotion.
复制标题
双微移液器技术在肿瘤细胞运动测量中的应用。
DOI：
10.1006/excr.1999.4388
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Dong,C
中科院分区：
文献类型：
--
作者：
You,J;Mastro,AM;Dong,C
这项工作的目的是表征肿瘤细胞运动的趋化刺激，使用双微量移液管测定。该测定涉及两个微量移液器。将单个A2058人黑素瘤细胞保留在约14 μm i.d.的移液管中，无压力梯度。将选择作为趋化源的IV型胶原蛋白溶液置于另一个移液管（10 μm o.d.）中，在移液管尖端处压力为零。然后将较小的移液管插入含有黑素瘤细胞的较大移液管中。初始化学引诱物浓度（C 0）和小移液管尖端与细胞表面之间的距离（δ）提供了肿瘤细胞朝向刺激运动的梯度（C 0/δ）。这种新的测定提供了细胞运动的直接测量：循环伪足突出（Lp）和随后的细胞运动（Lc）。研究了不同粘附基质对细胞运动的影响。Lp的峰长比最高运动速度（dLc/dt）提前一个明显的滞后时间，C 0/δ影响伪足伸出频率（fp）和dLc/dt，但对Lp影响不显著。基底粘附影响dLc/dt，但对Lporfp无明显影响。结论：伪足突出和基质粘附是肿瘤细胞运动的两个必要但相互独立的因素，dLc/dt与C 0/δ的变化相关，而C 0/δ与fp显著相关，与Lp无关。
The objective of this work was to characterize tumor cell locomotion in response to chemotactic stimulation using a dual-micropipet assay. The assay involves two micropipets. An individual A2058 human melanoma cell was retained, without pressure gradient, in a pipet of approximately 14 μm i.d. A solution of type IV collagen, chosen as the chemotactic source, was placed in another pipet (∼10 μm o.d.) with zero pressure at the pipet tip. The smaller pipet was then inserted into the larger one containing the melanoma cell. The initial chemoattractant concentration (C0) and the distance between the tip of the small pipet and the cell surface (δ) provided a gradient (C0/δ) for tumor cell locomotion toward stimulation. This novel assay provides a direct measure of cell movement: cyclic pseudopod protrusion (Lp) and subsequent cell locomotion (Lc). The influences of different adhesion substrates on cell locomotion were also studied. The peak length inLpprecedes the highest locomotion velocity (dLc/dt) by an apparent lag time.C0/δ influences pseudopod protrusion frequency (fp) anddLc/dt, but not significantly onLp. Substrate adhesions affectdLc/dt, but apparently notLporfp. In conclusion, pseudopod protrusion and substrate adhesion are two necessary but mutually independent factors in tumor cell locomotion.dLc/dtcorrelates with changes inC0/δ, which is in significant correlation withfpbut notLp.