Journal: Molecular Cancer Therapeutics

Article Title: Targeting of sonic hedgehog-GLI signaling: a potential strategy to improve therapy for mantle cell lymphoma

doi: 10.1158/1535-7163.mct-07-2118

Figure Lengend Snippet: Figure 1. Expression of Shh-GLI signaling components in MCL. A, schematic representation of Shh-GLI signaling in a cell. B, expression of Shh-GLI signaling molecules in MCL. The expression of transcripts associated with Shh-GLI signaling was determined in B cells from normal donors, primary MCL cells from patients, and JVM2, Granta 519, Jeko1, and Z138 human MCL cell lines by RT-PCR for PTCH, SMO, GLI1, and GLI2. Glyceraldehyde-3-phosphate dehydrogenase expression was used for normalization. PTCH, SMO, GLI1, and GLI2 genes were expressed in MCL cells, suggesting active Shh-GLI signaling in MCL. M, base pair standard. C, real-time PCR for Shh-GLI signaling molecules in MCL. Increased expression (lower Ct value) of PTCH, GLI1, and GLI2 in MCL cells compared with normal donor B cells suggests a role for Shh-GLI signaling in MCL. HPRT expression was used for normalization.

Article Snippet: Cells and Cell Culture Human MCL cell lines JVM2 (21) and Granta 519 and Jeko1 (22) were obtained from DSMZ, and Z138 cell line (21) was a gift from Dr. Michael Williams (University of Virginia).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: Molecular Cancer Therapeutics

Article Title: Targeting of sonic hedgehog-GLI signaling: a potential strategy to improve therapy for mantle cell lymphoma

doi: 10.1158/1535-7163.mct-07-2118

Figure Lengend Snippet: Figure 2. Perturbation of Shh-GLI signaling in MCL cells in vitro. The influence of exogenous Shh, with and without cyclopamine, was determined in the JVM2, Granta 519, Jeko1, and Z138 MCL cell lines at 24, 48, and 72 h by the [3H]thymidine uptake assay. The result was also corroborated by RT-PCR and real-time PCR for Shh-GLI signaling target transcript, GLI1, expression as shown in respective panels. A, JVM2. B, Granta 519. C, Jeko1. D, Z138. Perturbation of Shh-GLI signaling significantly influenced the proliferation of JVM2 MCL cells but did not influence the Granta 519, Jeko1, and Z138 MCL cells. Data for the experimental groups were calculated by considering the radioactive count obtained in cells cultured in control medium as 100%. Mean F SD from triplicates of at least three different experiments. Statistical analysis was done by the Student’s t test. RF2, RPMI containing 2% fetal bovine serum. Expression of glyceraldehyde-3-phosphate dehydrogenase and HPRT was used as housekeeping genes for RT-PCR and real-time PCR, respectively. Lower normalized Ct value represents the increased expression.

Article Snippet: Cells and Cell Culture Human MCL cell lines JVM2 (21) and Granta 519 and Jeko1 (22) were obtained from DSMZ, and Z138 cell line (21) was a gift from Dr. Michael Williams (University of Virginia).

Techniques: In Vitro, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction, Expressing, Cell Culture, Control

Journal: Molecular Cancer Therapeutics

Article Title: Targeting of sonic hedgehog-GLI signaling: a potential strategy to improve therapy for mantle cell lymphoma

doi: 10.1158/1535-7163.mct-07-2118

Figure Lengend Snippet: Figure 3. Influence of down-regulation of GLI in MCL cells in vitro on proliferation. Proliferation of MCL cells was determined in the presence of control medium, GLI1-ASO, GLI2-ASO, and control-ASO at 48, 72, and 96 h by the [3H]thymidine uptake assay. A, Jeko1. B, Z138. C, JVM2. D, Granta 519. There was a significant decrease in [3H]thymidine uptake in the presence of GLI1-ASO or GLI2-ASO compared with control cells or control-ASO, indicating that targeting of GLI will decrease MCL proliferation. Data for the experimental groups were calculated by considering the radioactive count in the cells cultured in control medium as 100%. Mean F SD from triplicates of at least three different experiments. Statistical analysis was done by the Student’s t test.

Article Snippet: Cells and Cell Culture Human MCL cell lines JVM2 (21) and Granta 519 and Jeko1 (22) were obtained from DSMZ, and Z138 cell line (21) was a gift from Dr. Michael Williams (University of Virginia).

Techniques: In Vitro, Control, Cell Culture

Journal: Molecular Cancer Therapeutics

Article Title: Targeting of sonic hedgehog-GLI signaling: a potential strategy to improve therapy for mantle cell lymphoma

doi: 10.1158/1535-7163.mct-07-2118

Figure Lengend Snippet: Figure 4. Influence of GLI-down-regulation on BCL2 and CCND1 expression in MCL. RT-PCR and real-time PCR for GLI1, GLI2, BCL2, and cyclin D1 were done using gene-specific primers, and glycer- aldehyde-3-phosphate dehydrogenase and HPRT were used for normalization in RT-PCR and real- time PCR, respectively. The levels of transcripts were decreased (higher Ct value) for cyclin D1 and BCL2 in the presence of both GLI1-ASO and GLI2- ASO compared with controls. A, JVM2. B, Granta 519. C, Jeko1.

Article Snippet: Cells and Cell Culture Human MCL cell lines JVM2 (21) and Granta 519 and Jeko1 (22) were obtained from DSMZ, and Z138 cell line (21) was a gift from Dr. Michael Williams (University of Virginia).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Real-time Polymerase Chain Reaction

Journal: Molecular Cancer Therapeutics

Article Title: Targeting of sonic hedgehog-GLI signaling: a potential strategy to improve therapy for mantle cell lymphoma

doi: 10.1158/1535-7163.mct-07-2118

Figure Lengend Snippet: Figure 5. Influence of GLI down- regulation on the susceptibility of MCL cells to chemotherapy in vitro. GLI- down-regulated MCL cells were cultured in the presence of doxorubicin for 24 h, and proliferation was determined by the [3H]thymidine uptake assay. Significant decreases in [3H]thymidine uptake in the presence of doxorubicin were noted when MCL cells were treated GLI1-ASO or GLI2-ASO compared with control doxorubicin or control-ASO, indicating GLI-mediated susceptibility of MCL to chemotherapy. A, Granta 519. B, JVM2. C, Jeko1. D, Z138. Data for the experimental groups were calculated by considering the radioactive count in the cells cultured in control medium as 100%. Mean F SD from triplicates of at least three different experiments. Statis- tical analysis was done by the Student’s t test. *, P < 0.05, compared with doxorubicin only; #, P < 0.05, com- pared with respective ASO only.

Article Snippet: Cells and Cell Culture Human MCL cell lines JVM2 (21) and Granta 519 and Jeko1 (22) were obtained from DSMZ, and Z138 cell line (21) was a gift from Dr. Michael Williams (University of Virginia).

Techniques: In Vitro, Cell Culture, Control