Single-Cell Proteomic Profiling Identifies Nanoparticle Enhanced Therapy for Triple Negative Breast Cancer Stem Cells.

Breast cancer remains a major cause of cancer-related deaths in women worldwide. Chemotherapy-promoted stemness and enhanced stem cell plasticity in breast cancer is a cause for great concern. The discovery of drugs targeting BCSCs was suggested to be an important advancement in the establishment of therapy that improves the efficacy of chemotherapy. In this work, by using single-cell mass cytometry, we observed that stemness in spheroid-forming cells derived from MDA-MB-231 cells was significantly increased after doxorubicin administration and up-regulated integrin αvß3 expression was also observed. An RGD-included nanoparticle (CS-V) was designed, and it was found that it could promote doxorubicin's efficacy against MDA-MB-231 spheroid cells. The above observations suggested that the combination of RGD-included nanoparticles (CS-V) with the chemo-drug doxorubicin could be developed as a potential therapy for breast cancer.

A pre-investigational new drug study of lung spheroid cell therapy for treating pulmonary fibrosis.

Idiopathic pulmonary fibrosis is a lethal interstitial lung disease with unknown etiology, no cure, and few treatment options. Herein, a therapy option is presented that makes use of a heterogeneous population of lung cells, including progenitor cells and supporting cells lines, cultured in adherent and suspension conditions, the latter of which induces spontaneous spheroid formation. Within these spheroids, progenitor marker expression is augmented. The cells, called lung spheroid cells, are isolated from fibrotic lungs, expanded, and delivered in single cell suspensions into rat models of pulmonary fibrosis via tail-vein injections. Two bleomycin-induced fibrotic rat models are used; a syngeneic Wistar-Kyoto rat model, treated with syngeneic cells, and a xenogeneic nude rat model, treated with human cells. The first objective was to study the differences in fibrotic progression in the two rat models after bleomycin injury. Nude rat fibrosis formed quickly and extended for 30 days with no self-resolution. Wistar-Kyoto rat fibrosis was more gradual and began to decrease in severity between days 14 and 30. The second goal was to find the minimum effective dose of cells that demonstrated safe and effective therapeutic value. The resultant minimum effective therapeutic dose, acquired from the nude rat model, was 3 × 106 human cells. Histological analysis revealed no evidence of tumorigenicity, increased local immunological activity in the lungs, or an increase in liver enzyme production. These data demonstrate the safety and efficacy of lung spheroid cells in their application as therapeutic agents for pulmonary fibrosis, as well as their potential for clinical translation.

Flat cells come full sphere: Are mutant cytoskeletal-related proteins oncoprotein-monsters or useful immunogens?

Osteogenesis imperfecta is inherited as a dominant disease because if one allele is mutated, it contributes a mutant, destructive subunit polypeptide to collagen, which requires many subunits to form normal, polymeric, collagenous structures. Recent cancer genome atlas (TCGA) data indicate that cytoskeletal-related proteins are among the most commonly mutated proteins in human cancers, in distinct mutation frequency groups, i.e., including low mutation frequency groups. Part of the explanation for this observation is likely to be the fact that many of the coding regions for these proteins are very large, and indeed, it is likely these coding regions are mutated in many cells that never become cancerous. However, it would not be surprising if mutations in cytoskeletal proteins, when combined with oncoprotein or tumor suppressor protein mutations, had significant impacts on cancer development, for a number of reasons, including results obtained almost 5 decades ago indicating that well-spread cells in tissue culture, with well-formed cytoskeletons, were less tumorigenic than spherical cells with disrupted cytoskeletons. This raises the question, are mutant cytoskeletal proteins, which would likely interfere with polymer formation, a new class of oncoproteins, in particular, dominant negative oncoproteins? If these proteins are so commonly mutant, could they be the bases for common cancer vaccines?

LY294002 decreases the proliferation of cancer stem cell-enriched spheroid cells from human hepatocel-lular carcinoma via inhibiting of PI3K-Akt signaling pathway / 国际肿瘤学杂志

Objective To investigate the impact of LY294002 on the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via regulating phosphatidylinositol-3-kinaseprotein kinase B(PI3K-Akt)signaling pathway. Methods The cancer stem cell-enriched spheroid cells were genera-ted by culturing HepG2 cells in serum-free medium. LY294002(10,20,30 μmol/ L),an inhibitor of PI3K-Akt signaling pathway,was used in the experimental groups,without used in the control group. The impact of LY294002 on the spheroid cells proliferation was confirmed by cell counting kit(CCK-8 kit). The expression of Akt was tested by Western blotting. The expression of PI3K-Akt signaling pathway downstream genes such as decoy receptor 3(DcR3),mammalian target of rapamycin(mTOR),B-cell lymphoma(Bcl)-2 and Cyclin D1 were tested by real-time PCR. Results 30 μmol/ L LY294002 could inhibit the proliferation of spheroid cells, and significant difference in the absorbance(A value)was observed between the experimental group and control group[(0. 14 ± 0. 03)vs(0. 56 ± 0. 01),t = - 8. 915,P = 0. 000]. The expression level of phosphorylated Akt protein increased[(0. 57 ± 0. 08)vs(0. 16 ± 0. 42),t = 6. 027,P = 0. 026]. The mRNA of DcR3 [(0. 38 ± 0. 08)vs 1,t = 13. 060,P = 0. 006],mTOR[(0. 37 ± 0. 04)vs 1,t = 30. 363,P = 0. 001],Bcl-2 [(0. 26 ± 0. 04)vs 1,t = 33. 554,P = 0. 001]and Cyclin D1[(0. 10 ± 0. 02)vs 1,t = 63. 528,P = 0. 000] decreased. Conclusion LY294002 could inhibit the proliferation of cancer stem cell-enriched spheroid cells from human hepatocellular carcinoma via inhibiting PI3K-Akt signaling pathway.

Biological properties of colon cancer spheroid cells cultured in serum-free medium / 中华消化外科杂志

Objective To obtain colon cancer spheroid cells from human colon cancer cell lines cultured in serum-free medium (SFM),and investigate the proliferative and migratory properties of colon cancer spheroid cells.Methods Human colon cancer cell lines HCT116 and HT29 were cultured in SFM,and then the generation of spheroid cells was observed.The expression of stem cell surface marker CD133 was detected by flow cytometry,and the proliferative and migratory properties of colon cancer spheroid cells were detected by cell counting kit-8 and Transwell migration assay,respectively.All data were analyzed by using the t test.Results Spheroid cells were obtained from colon cancer cell lines HCT116 and HT29 in SFM.The ratios of spheroid cells with positive expression of CD133 generated by HCT116 and HT29 were 75.44％ ± 11.41％ and 76.22％ ± 14.23％,respectively.Compared with original colon cancer cells cultured in serum supplemented medium,the number of HCT116 and HT29 spheroid cells with positive expression of CD133 was significantly greater (t =11.43,9.17,P ＜ 0.05 ),and the proliferative and migratory abilities were much stronger also.Conclusion Colon cancer spheroid cells cultured in SFM have higher positive expression of CD133 and stronger proliferative and migratory abilities,and it can be utilized as a feasible model for further studies of colonic stem cells.