methuosis是一種新的蛋白酶獨(dú)立細(xì)胞死亡的形式(編輯修改稿)
2024-10-06 00:54 本頁面
【文章內(nèi)容簡介】 mean 177。 SD from three separate cultures.Figure 3. Comparison of the abilities of MOMIPP and MIPP to induce the morphological hallmarks of methuosis. One day after plating, U251 GBM cells were treated with MOMIPP or MIPP at final concentrations of (A) or 10 μM (B). Controls received an equivalent volume of vehicle (DMSO). Cells were observed by phase contrast microscopy on three sequential days after addition of the pounds, without changing the medium or replenishing the pounds. Methuosis is characterized by extensive accumulation of phaselucent cytoplasmic vacuoles, with eventual cell rounding and detachment from the substratum as viability Figure 4. Comparison of the abilities of MOMIPP and MIPP to inhibit survival of U251 GBM cells in colonyforming assays. (A) Cells were plated for colonyforming assays as described in the Experimental Section. One day after plating the cells, MOMIPP (●) or MIPP (■) was added to the medium at the indicated concentrations, and cells were maintained in the presence of the pounds for 48 h. Thereafter, the pounds were removed, and colonies 50 cells were counted after 2 weeks. Each point represents the mean (177。SD) from three separate dishes, with the results expressed as percent of the mean of the parallel control dishes containing DMSO. (B) The effects MOMIPP (●) and MIPP (■) on colony formation were pared as in panel A, except that cells were exposed to the pounds for only 4 h instead of 48 h.為了確定誘導(dǎo)methuosis的化合物針對(duì)抗TMZ的神經(jīng)母細(xì)胞瘤是否有效，我們使用以前生成的基本上不受濃度高達(dá)100微米TMZ影響的抗TMZ膠質(zhì)瘤細(xì)胞系（10）（圖5A）如圖5b所示，MIPP和MOMIPP能夠誘導(dǎo)methuosis和降低抗TMZ細(xì)胞的細(xì)胞活性，但MOMIPP的效能明顯優(yōu)于MIPP。MOMIPP通過methuosis形式殺死有抗藥性的腫瘤細(xì)胞的能力并不僅僅局限于膠質(zhì)母細(xì)胞瘤。例如，我們觀察到類似情況，MOMIPP在親代和抗阿霉素的MCF7乳腺癌細(xì)胞（圖5C，D）引起空泡并導(dǎo)致其集落形成能力的降低。為了確定對(duì)腫瘤細(xì)胞造成最大限度毒性作用的濃度的MOMIPP是否也會(huì)對(duì)正常細(xì)胞產(chǎn)生毒性，我們用10μMMOMIPP處理人類乳腺上皮細(xì)胞（HMEC）。圖5E所示，在加入MOMIPP后24小時(shí)內(nèi)HMECs進(jìn)行大范圍的細(xì)胞質(zhì)空泡化。到48小時(shí)，細(xì)胞活力減少約40％，而在類似的細(xì)胞密度下同期處理的MCF7乳腺癌細(xì)胞減少了70％（圖5F）。為了進(jìn)一步評(píng)估MOMIPP對(duì)正常細(xì)胞的影響，我們把化合物應(yīng)用到人體皮膚成纖維細(xì)胞。在所有其他細(xì)胞測(cè)試實(shí)驗(yàn)中，到24小時(shí)10μMMOMIPP明顯誘導(dǎo)成纖維細(xì)胞空泡化（圖5G）。與HMECs類似，接種在合適密度來保持指數(shù)增長的正常成纖維細(xì)胞在接觸到MOMIPP 48小時(shí)時(shí)，表現(xiàn)出生存能力下降40％的特點(diǎn)。然而，當(dāng)纖維母細(xì)胞接種在一個(gè)較高的初始密度，使他們?cè)贛OMIPP加入時(shí)以形成固定相，那么細(xì)胞活性基本上不受影響（圖5H），即使細(xì)胞發(fā)生空泡（未顯示）。這些觀察表明，雖然MOMIPP會(huì)對(duì)正常細(xì)胞產(chǎn)生毒性，但與癌癥細(xì)胞系，如U251和MCF7相比，正常細(xì)胞對(duì)這類化合物相對(duì)不太敏感。MOMIPP對(duì)亞融合與融合的成纖維細(xì)胞的影響差別提供了一個(gè)可能的解釋，這意味著，以空泡為典型的內(nèi)體販運(yùn)的中斷可能在不分裂的細(xì)胞中比在正常增殖的細(xì)胞中具有相對(duì)較好的耐受性。 Figure 5. MOMIPP effectively inhibits the viability of drugresistant GBM and breast cancer cells. (A) TMZresistant U251 glioblastoma cells (U251TR) were derived as described previously.(10) The graph, reprinted from ref 10 with permission, shows that in contrast to the parental U251 cells, the viability of U251TR cells is not reduced by treatment with TMZ. (B) The U251TR cells were treated with the indicated concentrations of MIPP or MOMIPP for 48 h and then subjected to colonyforming assays as described in the Experimental Section. Each point is based on colony counts from three dishes (mean 177。 SD), with the results expressed as percent of the mean from parallel control dishes treated with an equivalent volume of vehicle alone (DMSO). (C) Parental and doxorubicinresistant (DoxR) MCF7 breast cancer cells(56) were treated with 10 μM MOMIPP for 24 h and then examined by phasecontrast microscopy. (D) Longterm viability of parental or DoxR MCF7 cells was assessed by colonyforming assay after a 48 h of treatment with the indicated concentrations of MOMIPP. The results are the mean 177。 SD of determinations performed on three parallel dishes. (E) Normal HMECs were treated with 10 μM MOMIPP or an equivalent volume of DMSO (control) and examined by phasecontrast microscopy after 24 h. (F) MCF7 cells or HMECs were plated in 96well plates. After 48 h, while the cells were still subconfluent, fr
環(huán)評(píng)公示相關(guān)推薦
鄂ICP備17016276號(hào)-1