human brain cancer stem cell bcsc line  (Celprogen Inc)

Journal: Naunyn-Schmiedeberg's Archives of Pharmacology

Article Title: Anti-miRNA- 155 - 5p enhances the cytotoxic efficacy of temozolomide in brain cancer stem cells

doi: 10.1007/s00210-025-04135-6

Figure Lengend Snippet: Expression levels of miR- 155 - 5p. a ) Expression levels in glioblastoma and healthy brain tissues (To compare gene expression levels across different samples regardless of sequencing depth, Reads Per Million (RPM) is calculated by dividing the raw read count of a given transcript by the total number of mapped reads in the sample and multiplying by 10 6 .); b ) Expression levels in BCSCs compared to BSCs (RPM: Reads per million); c) Expression levels in anti-miR- 155 - 5p treated BCSCs and BSCs compared to untreated control cells Statistical significance was evaluated by Student’s t-test ( p < 0.05: *, p < 0.01: **, p < 0.001: ***, p < 0.0001: ****)

Article Snippet: BCSCs were obtained from Celprogen (Catalog No: 36110–37), and BSCs were sourced from the same provider (Catalog No: 36109–36, USA).

Techniques: Expressing, Gene Expression, Sequencing, Control

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: miR-34a is the most pronounced microRNA for further research in long-term-cultured BCSC. ( A ) Capacities of self-renewal and sphere-formation of XM322 (left panel) and XM607 (right panel). Scale bar, 100 µm. ( B ) Capacities of self-renewal and sphere-formation of MDA-MB-231.SC (left panel) and MCF-7.SC (right panel). Scale bar, 100 µm. ( C ) Heatmap presents microRNA expressions among immortalized healthy mammary epithelial cells (HME, 184A1 and MCF-12A), breast cancer cells (BC, contained MDA-MB-231, MCF-7, MDA-MB-468 and SK-BR-3), and BCSC (contained MDA-MB-231.SC, MCF-7.SC, XM322 and XM607). ( D ) qRT-PCR analysis indicated that miR-34a expression of HME is significantly higher than that of BC and BCSC. miR-34a expression was normalized to the endogenous control 5S rRNA. NS: not significant. ( E ) qRT-PCR analyses indicated that samples within the stage I-II group had significantly lower miR-34a expression compared with those in groups of the adjacent healthy breast tissue specimens (HBT); additionally, miR-34a expression within stage III patients was lowest among the three groups. ***p<0.001.

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: Cell Culture, Quantitative RT-PCR, Expressing

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: TV-miR-34a induces high throughput of miR-34a expression in BCSC. ( A ) Expression of hTERT was rarely detectable in both HME cells; while hTERT was highly expressed in all BC and BCSC. ( B ) Telomerase activities in HME cells were lower than BCs and BCSC. NS: not significant. ( C ) Representative images of isolated single cell (upper panel) and pair cells (low panel) expressing GFP-labeled TV-miR-34a in XM322. Scale bar, 5 µm ( D ) Telomerase activities among HME, BC and BCSC did not alter significantly following their corresponding transfection with Ctrl, TV-miR-Ctrl or TV-miR-34a, respectively. Scale bar, 5 µm.( E ) qRT-PCR analysis demonstrated that TV-miR-34a plasmid significantly up-regulated miR-34a expression comparing with either TV-miR-Ctrl or Ctrl in both BCs and BCSC. ( F ) MTT assays of cell viabilities showed that TV-miR-34a decreased the cell viabilities in both BCSC (MDA-MB-468, SK-BR-3, MDA-MB-231, MCF-7, parental cells of XM322 and XM607) and BCs (MDA-MB-231.SC, MCF-7.SC, XM322 and XM607); whereas, cell viabilities of HME (MCF-12A and 184A1) were not found to be significantly different. The inhibitory effect of TV-miR-34a nanopartiple on cell viabilities of BCSC was stronger than their corresponding parental breast cancer cells. *p<0.05, **p<0.01, ***p<0.001

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: High Throughput Screening Assay, Expressing, Isolation, Labeling, Transfection, Quantitative RT-PCR, Plasmid Preparation

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: TV-miR-34a inhibits tumor-initiating properties of long-term-cultured BCSC in vitro . ( A ) Mammospheres of XM607 initiated adherence and differentiation following TV-miR-34a interference. Scale bar, 100 µm. ( B ) Representative images (left panel) and statistical results (right panel) of TV-miR-34a robustly and persistently decreasing the population of CD44 + CD24 - XM607; while miR-34a influence remained transient and reversible. ( C ) Representative images (left panel; scale bar, 100 µm) and statistical result (right panel) of clonogenesis abilities of MCF-7.SC down-regulated by TV-miR-34a. ***p<0.001.

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: Cell Culture, In Vitro

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: TV-miR-34a plays an inhibitory effect on tumor-initiating properties of long-term-cultured BCSC via directly targeting C22ORF28. ( A ) Schematic diagram of Luc-C22ORF28-3'UTR and mutation of Luc-C22ORF28-3'UTR (Luc-C22ORF28-3'UTR-mut) for presence of miR-34a conversed binding sites. ( B ) Mutating the predicted miR-34a binding sites within the Luc-C22ORF28-3'UTR luciferase reporter significantly abolished TV-miR-34a-dependent repression. ( C ) LIN28A is rarely detected in BCSC. Both C22ORF28 and CD44, rather than LIN28A, were the inhibitory targets of TV-miR-34a treatment. ( D ) Constructions of C22ORF28-Ad and C22ORF28-KD can effectively up-regulate and down-regulate expressions of C22ORF28 and CD44 in BCSC, respectively. ( E ) C22ORF28-Ad increased percentage of CD44 + CD24 - subpopulation cells in BCSC, whereas C22ORF28-KD decreased these subpopulation cells. ( F ) Representative images (left panel) and statistical results (right panel) of C22ORF28-Ad promoting sphere-formation ability within soft agarose of XM322, but C22ORF28-KD eliminating this ability. Scale bar, 100 µm. ( G ) Representative images (left panel) and statistical result (right panel) of TV-miR-34a suppressing CD44 expression measured by clonal pair-cell analyses in XM607. Scale bar, 10 µm; C.E.: co-expressed; M.E.: mutually exclusive. Each experiment was repeated at least three times. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: Cell Culture, Mutagenesis, Binding Assay, Luciferase, Expressing

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: Evaluation of the inhibitory effect of C22ORF28 intervene on long-term-cultured BCSC during TV-miR-34a treatment. ( A ) Representative images of mammosphere formation and fluorescence expression of GFP-labeled BCSC following transduction with Ctrl, TV-miR-Ctrl, TV-miR-34a, co-transfection of TV-miR-34a plus C22ORF28-3'-UTR, as well as co-transfection of TV-miR-34a plus C22ORF28-3'-UTR-mut, respectively (L.L.: lamp light; E.L.: exciting light). Scale bar, 100 µm. ( B ) Statistical results of differences in mammosphere formation. ( C ) Statistical results of fluorescence intensity. Each experiment was repeated at least in triplicate. ( D ) Schematic diagram of C22ORF28 mediating TV-miR-34a treatment in GFP-labeled long-term-cultured BCSC. **p<0.01, ***p<0.001.

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: Cell Culture, Fluorescence, Expressing, Labeling, Transduction, Cotransfection

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: TV-miR-34a has synergistic effects with docetaxel in eradication of BCSC. ( A ) MDA-MB-231.SC was observed with lamp light (L.L., left panel) or exciting light (E.L., right panel) to detect the depression effects. Scale bar, 100 µm. ( B ) Statistical results of differences in fluorescence expressions among groups of Ctrl, TV-miR-34a, and presence or absence of docetaxel 5 days following TV-miR-34a transduction. ( C ) MTT assay. ( D ) Docetaxel alone did not significantly alter expressions of CD44 and C22ORF28 in BCSC, compared to untreated cells. Combined application of TV-miR-34a plus docetaxel on BCSC for 48 h significantly decreased expressions of CD44 and C22ORF28 compared with TV-miR-34a alone. Cell line-derived (upper panel); tumor tissue-derived BCSC (low panel). ( E ) Schematic Diagram. All data corresponds to the mean ± SD of three independent experiments.. *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: Fluorescence, Transduction, MTT Assay, Derivative Assay

Journal: Theranostics

Article Title: Nanoparticle Delivery of miR-34a Eradicates Long-term-cultured Breast Cancer Stem Cells via Targeting C22ORF28 Directly

doi: 10.7150/thno.20771

Figure Lengend Snippet: Schematic Diagram of the TV-miR-34a System. ( A ) The hTERT promoter drives expression of the GAL4-VP2 fusion protein selectively in BCSC, but not in adult stem cells ( B ) GAL4-VP2 is expressed; ( C ) GAL4-VP2 discerns the promoter G5E4T; ( D ) G5E4T promotes the expression of the miR-34a protein; ( E ) the mature miR-34a are incorporated into RISC, which inhibits C22ORF28 expression leading to eradication of BCSC. ( F ) Telomeres (mainly composed by hTERT) form G-quadruplex structures (G4), which along with G4 can cause replication fork stalling.

Article Snippet: To determine the optimum antitumor dose of T-VISA-miR-34a plasmid in vivo , a suspension of luciferase-labeled BCSC (1×10 4 cells) was inoculated at the left fourth inguinal mammary gland of female BALB/c-nude mice (6-week-old; Vital River Laboratories Animal, Beijing, China).

Techniques: Expressing