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Tetrandrine induces autophagy and differentiation by activating ROS and Notch1 signaling in leukemia cells.

Liu T, Men Q, Wu G, Yu C, Huang Z, Liu X, Li W - Oncotarget (2015)

Bottom Line: Tetrandrine is a traditional Chinese medicinal herb extract with antitumor effects.The in vivo results indicated that low concentrations of tetrandrine inhibited leukemia cells proliferation and induced autophagy and then facilitated their differentiation, by activating ROS and Notch1 signaling.We suggest that tetrandrine is a potential agent for the treatment of APL by inducing differentiation of leukemia cells.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Wuhan University, Wuhan, P. R. China.

ABSTRACT
All-trans retinoic acid (ATRA) is a differentiating agent for the treatment of acute promyelocytic leukemia (APL). However, the therapeutic efficacy of ATRA has limitations. Tetrandrine is a traditional Chinese medicinal herb extract with antitumor effects. In this study, we investigated the effects of tetrandrine on human PML-RARα-positive acute promyelocytic leukemia cells. Tetrandrine inhibited tumors in vivo. It induced autophagy and differentiation by triggering ROS generation and activating Notch1 signaling. Tetrandrine induced autophagy and differentiation in M5 type patient primary leukemia cells. The in vivo results indicated that low concentrations of tetrandrine inhibited leukemia cells proliferation and induced autophagy and then facilitated their differentiation, by activating ROS and Notch1 signaling. We suggest that tetrandrine is a potential agent for the treatment of APL by inducing differentiation of leukemia cells.

No MeSH data available.


Related in: MedlinePlus

Tetrandrine facilitates NB4 cell differentiationOxidant dimethylsulfoxide (DMSO) was used as a negative control (Con), and All-trans retinoic acid (RA) treatment was used as a positive control. (A) Cell morphology was observed under a microscope with or without Wright–Giemsa staining after 4 days of 2 μM tetrandrine (Tet) treatment. Data are represented as mean ± SD. **p <0.01. (B) CD11b and CD14 antigens expression was measured alone (C) or co-measured by flow cytometry after 4 days of tetrandrine (Tet) treatment. (D) The respective cell surface expression of the CD11b and CD14 antigens, on NB4 cells after tetrandrine (Tet) treatment at the indicated concentrations and times was measured by flow cytometry. (E) NBT reduction assay analysis of differentiation. NB4 cells treated with 2 μM tetrandrine (Tet) for 24h-96h. Data are represented as mean ± SD. **p <0.01. (F) The relative levels of MMP9 expression were measured by RT-PCR after 2 μM tetrandrine (Tet) treatment for 24h-96h. Data are represented as mean ±SD. *p <0.05.
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Figure 5: Tetrandrine facilitates NB4 cell differentiationOxidant dimethylsulfoxide (DMSO) was used as a negative control (Con), and All-trans retinoic acid (RA) treatment was used as a positive control. (A) Cell morphology was observed under a microscope with or without Wright–Giemsa staining after 4 days of 2 μM tetrandrine (Tet) treatment. Data are represented as mean ± SD. **p <0.01. (B) CD11b and CD14 antigens expression was measured alone (C) or co-measured by flow cytometry after 4 days of tetrandrine (Tet) treatment. (D) The respective cell surface expression of the CD11b and CD14 antigens, on NB4 cells after tetrandrine (Tet) treatment at the indicated concentrations and times was measured by flow cytometry. (E) NBT reduction assay analysis of differentiation. NB4 cells treated with 2 μM tetrandrine (Tet) for 24h-96h. Data are represented as mean ± SD. **p <0.01. (F) The relative levels of MMP9 expression were measured by RT-PCR after 2 μM tetrandrine (Tet) treatment for 24h-96h. Data are represented as mean ±SD. *p <0.05.

Mentions: The strategy of inducing the differentiation of leukemia cells is significant for APL clinical treatment. To examine whether tetrandrine could stimulate the differentiation of NB4 cells, we first evaluated cellular maturation by observing the cellular morphology with Wright-Giemsa staining. As shown in Fig. 5A, NB4 cells treated with tetrandrine had the typical characteristic morphology of differentiated cells, such as an irregular nucleus, the presence of vacuoles and a large nuclear/cytoplasm ratio. Next, we further predicted NB4 cell differentiation by analyzing the expression of the cell surface differentiation-related antigens, CD14 and CD11b. The results revealed that 2 μM tetrandrine enhanced CD14 and CD11b expression on the NB4 cell surface, either alone or in combination, as shown by immunofluorescence labeling of these two proteins. In contrast, 1 μM tetrandrine did not significantly induce NB4 cell differentiation (Fig. 5B, C and D). The NBT reduction assay, which is a classic experiment to detect cellular differentiation, also found that NBT reduction activity increased remarkably with tetrandrine treatment and reached a peak value at 48 hours (Fig. 5E). MMP9 levels can be used as an additional NB4 cell differentiation marker. Here, we also observed that tetrandrine treatment promoted a significant increase of MMP9 mRNA levels (Fig. 5F). Thus, these results suggest that 2 μM tetrandrine promoted NB4 cell differentiation.


Tetrandrine induces autophagy and differentiation by activating ROS and Notch1 signaling in leukemia cells.

Liu T, Men Q, Wu G, Yu C, Huang Z, Liu X, Li W - Oncotarget (2015)

Tetrandrine facilitates NB4 cell differentiationOxidant dimethylsulfoxide (DMSO) was used as a negative control (Con), and All-trans retinoic acid (RA) treatment was used as a positive control. (A) Cell morphology was observed under a microscope with or without Wright–Giemsa staining after 4 days of 2 μM tetrandrine (Tet) treatment. Data are represented as mean ± SD. **p <0.01. (B) CD11b and CD14 antigens expression was measured alone (C) or co-measured by flow cytometry after 4 days of tetrandrine (Tet) treatment. (D) The respective cell surface expression of the CD11b and CD14 antigens, on NB4 cells after tetrandrine (Tet) treatment at the indicated concentrations and times was measured by flow cytometry. (E) NBT reduction assay analysis of differentiation. NB4 cells treated with 2 μM tetrandrine (Tet) for 24h-96h. Data are represented as mean ± SD. **p <0.01. (F) The relative levels of MMP9 expression were measured by RT-PCR after 2 μM tetrandrine (Tet) treatment for 24h-96h. Data are represented as mean ±SD. *p <0.05.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4480730&req=5

Figure 5: Tetrandrine facilitates NB4 cell differentiationOxidant dimethylsulfoxide (DMSO) was used as a negative control (Con), and All-trans retinoic acid (RA) treatment was used as a positive control. (A) Cell morphology was observed under a microscope with or without Wright–Giemsa staining after 4 days of 2 μM tetrandrine (Tet) treatment. Data are represented as mean ± SD. **p <0.01. (B) CD11b and CD14 antigens expression was measured alone (C) or co-measured by flow cytometry after 4 days of tetrandrine (Tet) treatment. (D) The respective cell surface expression of the CD11b and CD14 antigens, on NB4 cells after tetrandrine (Tet) treatment at the indicated concentrations and times was measured by flow cytometry. (E) NBT reduction assay analysis of differentiation. NB4 cells treated with 2 μM tetrandrine (Tet) for 24h-96h. Data are represented as mean ± SD. **p <0.01. (F) The relative levels of MMP9 expression were measured by RT-PCR after 2 μM tetrandrine (Tet) treatment for 24h-96h. Data are represented as mean ±SD. *p <0.05.
Mentions: The strategy of inducing the differentiation of leukemia cells is significant for APL clinical treatment. To examine whether tetrandrine could stimulate the differentiation of NB4 cells, we first evaluated cellular maturation by observing the cellular morphology with Wright-Giemsa staining. As shown in Fig. 5A, NB4 cells treated with tetrandrine had the typical characteristic morphology of differentiated cells, such as an irregular nucleus, the presence of vacuoles and a large nuclear/cytoplasm ratio. Next, we further predicted NB4 cell differentiation by analyzing the expression of the cell surface differentiation-related antigens, CD14 and CD11b. The results revealed that 2 μM tetrandrine enhanced CD14 and CD11b expression on the NB4 cell surface, either alone or in combination, as shown by immunofluorescence labeling of these two proteins. In contrast, 1 μM tetrandrine did not significantly induce NB4 cell differentiation (Fig. 5B, C and D). The NBT reduction assay, which is a classic experiment to detect cellular differentiation, also found that NBT reduction activity increased remarkably with tetrandrine treatment and reached a peak value at 48 hours (Fig. 5E). MMP9 levels can be used as an additional NB4 cell differentiation marker. Here, we also observed that tetrandrine treatment promoted a significant increase of MMP9 mRNA levels (Fig. 5F). Thus, these results suggest that 2 μM tetrandrine promoted NB4 cell differentiation.

Bottom Line: Tetrandrine is a traditional Chinese medicinal herb extract with antitumor effects.The in vivo results indicated that low concentrations of tetrandrine inhibited leukemia cells proliferation and induced autophagy and then facilitated their differentiation, by activating ROS and Notch1 signaling.We suggest that tetrandrine is a potential agent for the treatment of APL by inducing differentiation of leukemia cells.

View Article: PubMed Central - PubMed

Affiliation: College of Life Sciences, Wuhan University, Wuhan, P. R. China.

ABSTRACT
All-trans retinoic acid (ATRA) is a differentiating agent for the treatment of acute promyelocytic leukemia (APL). However, the therapeutic efficacy of ATRA has limitations. Tetrandrine is a traditional Chinese medicinal herb extract with antitumor effects. In this study, we investigated the effects of tetrandrine on human PML-RARα-positive acute promyelocytic leukemia cells. Tetrandrine inhibited tumors in vivo. It induced autophagy and differentiation by triggering ROS generation and activating Notch1 signaling. Tetrandrine induced autophagy and differentiation in M5 type patient primary leukemia cells. The in vivo results indicated that low concentrations of tetrandrine inhibited leukemia cells proliferation and induced autophagy and then facilitated their differentiation, by activating ROS and Notch1 signaling. We suggest that tetrandrine is a potential agent for the treatment of APL by inducing differentiation of leukemia cells.

No MeSH data available.


Related in: MedlinePlus