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B7-H3 silencing by RNAi inhibits tumor progression and enhances chemosensitivity in U937 cells.

Zhang W, Wang J, Wang Y, Dong F, Zhu M, Wan W, Li H, Wu F, Yan X, Ke X - Onco Targets Ther (2015)

Bottom Line: Downregulation of B7-H3 significantly decreased U937 cell growth and colony-forming ability.The cell migration rate of B7-H3 knockdown cells was reduced more than fivefold, and invasion capacity decreased by 86.7%.B7-H3 RNAi profoundly increased the antitumor effect of chemotherapy in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology and Lymphoma Research Center, Peking University Third Hospital, Beijing, People's Republic of China.

ABSTRACT

Background: The role of B7-H3 in acute monocytic leukemia U937 cells has not been thoroughly investigated.

Materials and methods: B7-H3 knockdown in the U937 cell line was performed using small hairpin (sh)RNA lentivirus transduction. The effects on cell proliferation, cycle, migration, and invasion were investigated by Cell Counting Kit-8 assay, methyl cellulose colony-forming assay, propidium iodide staining, and Transwell assays in vitro. Changes in cell growth inhibition and apoptosis, when combined with chemotherapy drugs, were determined using the Cell Counting Kit-8 and Annexin V-FITC/PI assays. U937 xenograft models were used to assess the effects of B7-H3 on tumorigenicity and the therapeutic effect of B7-H3 knockdown in combination with chemotherapy drugs in vivo.

Results: Downregulation of B7-H3 significantly decreased U937 cell growth and colony-forming ability. The mean inhibition rate of tumor growth with B7-H3 knockdown was 59.4%, and the expression of both Ki-67 and PCNA in xenografts was significantly reduced. After B7-H3 silencing, the U937 cell cycle was arrested at the G0/G1 phase. The cell migration rate of B7-H3 knockdown cells was reduced more than fivefold, and invasion capacity decreased by 86.7%. B7-H3 RNAi profoundly increased the antitumor effect of chemotherapy in vitro and in vivo. On day 19, inhibition rates of tumor growth in B7-H3 shRNA combined with idarubicin, cytarabine, and idarubicin plus cytarabine were 70.5%, 80.0%, and 90.0%, respectively (P=0.006, P=0.004, and P=0.016, respectively).

Conclusion: B7-H3 may promote U937 cell progression, and shRNA targeting B7-H3 significantly enhances sensitivity to chemotherapeutic drugs. These results may provide new insight into the function of B7-H3 and a promising therapeutic approach targeting B7-H3 in acute monocytic leukemia.

No MeSH data available.


Related in: MedlinePlus

B7-H3 knockdown increases chemosensitivity in vivo.Notes: (A) pKD combined with IDA and/or Ara-C reduced the tumor growth more effectively than in the pNC combined with chemotherapy groups, respectively. (B) The inhibition rates of tumor growth in the B7-H3 shRNA combined with chemotherapy groups were significantly higher than in the control plasmid combined with chemotherapy groups. (C) Excised xenograft tumors on day 19. (D) Mouse weight in ten different groups labeled 1)-10) was measured on day 7 and 19, respectively. All of the mice treated with B7-H3 shRNA exhibited no significant body weight loss compared with the relative pNC control groups on day 19. The label 1)-10) represents the ten groups of the experiment to detect the effect of B7-H3 RNAi on chemosensitivity in vivo. *Indicates that the results are statistically significant as compared to relative pNC when combining the chemotherapy groups; P<0.05.Abbreviations: NS, normal saline; IDA, idarubicin; pNC, nontargeted control plasmid; pKD, B7-H3 small hairpin RNA plasmid; d, days; Ara-C, cytarabine; shRNA, small hairpin RNA.
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f6-ott-8-1721: B7-H3 knockdown increases chemosensitivity in vivo.Notes: (A) pKD combined with IDA and/or Ara-C reduced the tumor growth more effectively than in the pNC combined with chemotherapy groups, respectively. (B) The inhibition rates of tumor growth in the B7-H3 shRNA combined with chemotherapy groups were significantly higher than in the control plasmid combined with chemotherapy groups. (C) Excised xenograft tumors on day 19. (D) Mouse weight in ten different groups labeled 1)-10) was measured on day 7 and 19, respectively. All of the mice treated with B7-H3 shRNA exhibited no significant body weight loss compared with the relative pNC control groups on day 19. The label 1)-10) represents the ten groups of the experiment to detect the effect of B7-H3 RNAi on chemosensitivity in vivo. *Indicates that the results are statistically significant as compared to relative pNC when combining the chemotherapy groups; P<0.05.Abbreviations: NS, normal saline; IDA, idarubicin; pNC, nontargeted control plasmid; pKD, B7-H3 small hairpin RNA plasmid; d, days; Ara-C, cytarabine; shRNA, small hairpin RNA.

Mentions: In order to explore the impact of B7-H3 knockdown on the antitumor activity of chemotherapy drugs in vivo, we constructed a tumor-bearing mouse model injected with nontransfected U937 cells. The treatments began on day 7, when the average tumor volumes reached 100 mm3. As shown in Figure 6A, the plasmid of B7-H3 shRNA (pKD) combined with IDA and/or Ara-C was more effective in reducing the established U937 tumor growth compared to the pNC groups combined with chemotherapy, while there were no significant differences in the tumor volumes between the pNC combined with chemotherapy groups and the chemotherapy groups alone. On day 19, the rates of tumor growth inhibition in B7-H3 shRNA combined with the IDA, Ara-C, and IDA + Ara-C groups were 70.5%, 80.0%, and 90.0%, respectively, which were higher than in the relative control plasmid combined with chemotherapy groups at 39.2%, 44.0%, and 73.9% (P=0.006, P=0.004, and P=0.016, respectively) (Figure 6B and C). The B7-H3 shRNA combined with IDA (3 mg/kg) and Ara-C (30 mg/kg) treatment group exhibited the best antitumor activity. Furthermore, all mice treated with B7-H3 shRNA (group 4),7), and 10) of ten tumor-bearing mice groups, as described earlier) exhibited no body weight loss at the end of our experiment on day 19 compared with the mice treated with pNC treatments (group 3), 6), and 9) of ten tumor-bearing mice groups, as described earlier) (Figure 6D). These results indicate that B7-H3 silencing can apparently enhance the chemosensitivity of U937 cells in a xenograft model.


B7-H3 silencing by RNAi inhibits tumor progression and enhances chemosensitivity in U937 cells.

Zhang W, Wang J, Wang Y, Dong F, Zhu M, Wan W, Li H, Wu F, Yan X, Ke X - Onco Targets Ther (2015)

B7-H3 knockdown increases chemosensitivity in vivo.Notes: (A) pKD combined with IDA and/or Ara-C reduced the tumor growth more effectively than in the pNC combined with chemotherapy groups, respectively. (B) The inhibition rates of tumor growth in the B7-H3 shRNA combined with chemotherapy groups were significantly higher than in the control plasmid combined with chemotherapy groups. (C) Excised xenograft tumors on day 19. (D) Mouse weight in ten different groups labeled 1)-10) was measured on day 7 and 19, respectively. All of the mice treated with B7-H3 shRNA exhibited no significant body weight loss compared with the relative pNC control groups on day 19. The label 1)-10) represents the ten groups of the experiment to detect the effect of B7-H3 RNAi on chemosensitivity in vivo. *Indicates that the results are statistically significant as compared to relative pNC when combining the chemotherapy groups; P<0.05.Abbreviations: NS, normal saline; IDA, idarubicin; pNC, nontargeted control plasmid; pKD, B7-H3 small hairpin RNA plasmid; d, days; Ara-C, cytarabine; shRNA, small hairpin RNA.
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Related In: Results  -  Collection

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f6-ott-8-1721: B7-H3 knockdown increases chemosensitivity in vivo.Notes: (A) pKD combined with IDA and/or Ara-C reduced the tumor growth more effectively than in the pNC combined with chemotherapy groups, respectively. (B) The inhibition rates of tumor growth in the B7-H3 shRNA combined with chemotherapy groups were significantly higher than in the control plasmid combined with chemotherapy groups. (C) Excised xenograft tumors on day 19. (D) Mouse weight in ten different groups labeled 1)-10) was measured on day 7 and 19, respectively. All of the mice treated with B7-H3 shRNA exhibited no significant body weight loss compared with the relative pNC control groups on day 19. The label 1)-10) represents the ten groups of the experiment to detect the effect of B7-H3 RNAi on chemosensitivity in vivo. *Indicates that the results are statistically significant as compared to relative pNC when combining the chemotherapy groups; P<0.05.Abbreviations: NS, normal saline; IDA, idarubicin; pNC, nontargeted control plasmid; pKD, B7-H3 small hairpin RNA plasmid; d, days; Ara-C, cytarabine; shRNA, small hairpin RNA.
Mentions: In order to explore the impact of B7-H3 knockdown on the antitumor activity of chemotherapy drugs in vivo, we constructed a tumor-bearing mouse model injected with nontransfected U937 cells. The treatments began on day 7, when the average tumor volumes reached 100 mm3. As shown in Figure 6A, the plasmid of B7-H3 shRNA (pKD) combined with IDA and/or Ara-C was more effective in reducing the established U937 tumor growth compared to the pNC groups combined with chemotherapy, while there were no significant differences in the tumor volumes between the pNC combined with chemotherapy groups and the chemotherapy groups alone. On day 19, the rates of tumor growth inhibition in B7-H3 shRNA combined with the IDA, Ara-C, and IDA + Ara-C groups were 70.5%, 80.0%, and 90.0%, respectively, which were higher than in the relative control plasmid combined with chemotherapy groups at 39.2%, 44.0%, and 73.9% (P=0.006, P=0.004, and P=0.016, respectively) (Figure 6B and C). The B7-H3 shRNA combined with IDA (3 mg/kg) and Ara-C (30 mg/kg) treatment group exhibited the best antitumor activity. Furthermore, all mice treated with B7-H3 shRNA (group 4),7), and 10) of ten tumor-bearing mice groups, as described earlier) exhibited no body weight loss at the end of our experiment on day 19 compared with the mice treated with pNC treatments (group 3), 6), and 9) of ten tumor-bearing mice groups, as described earlier) (Figure 6D). These results indicate that B7-H3 silencing can apparently enhance the chemosensitivity of U937 cells in a xenograft model.

Bottom Line: Downregulation of B7-H3 significantly decreased U937 cell growth and colony-forming ability.The cell migration rate of B7-H3 knockdown cells was reduced more than fivefold, and invasion capacity decreased by 86.7%.B7-H3 RNAi profoundly increased the antitumor effect of chemotherapy in vitro and in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology and Lymphoma Research Center, Peking University Third Hospital, Beijing, People's Republic of China.

ABSTRACT

Background: The role of B7-H3 in acute monocytic leukemia U937 cells has not been thoroughly investigated.

Materials and methods: B7-H3 knockdown in the U937 cell line was performed using small hairpin (sh)RNA lentivirus transduction. The effects on cell proliferation, cycle, migration, and invasion were investigated by Cell Counting Kit-8 assay, methyl cellulose colony-forming assay, propidium iodide staining, and Transwell assays in vitro. Changes in cell growth inhibition and apoptosis, when combined with chemotherapy drugs, were determined using the Cell Counting Kit-8 and Annexin V-FITC/PI assays. U937 xenograft models were used to assess the effects of B7-H3 on tumorigenicity and the therapeutic effect of B7-H3 knockdown in combination with chemotherapy drugs in vivo.

Results: Downregulation of B7-H3 significantly decreased U937 cell growth and colony-forming ability. The mean inhibition rate of tumor growth with B7-H3 knockdown was 59.4%, and the expression of both Ki-67 and PCNA in xenografts was significantly reduced. After B7-H3 silencing, the U937 cell cycle was arrested at the G0/G1 phase. The cell migration rate of B7-H3 knockdown cells was reduced more than fivefold, and invasion capacity decreased by 86.7%. B7-H3 RNAi profoundly increased the antitumor effect of chemotherapy in vitro and in vivo. On day 19, inhibition rates of tumor growth in B7-H3 shRNA combined with idarubicin, cytarabine, and idarubicin plus cytarabine were 70.5%, 80.0%, and 90.0%, respectively (P=0.006, P=0.004, and P=0.016, respectively).

Conclusion: B7-H3 may promote U937 cell progression, and shRNA targeting B7-H3 significantly enhances sensitivity to chemotherapeutic drugs. These results may provide new insight into the function of B7-H3 and a promising therapeutic approach targeting B7-H3 in acute monocytic leukemia.

No MeSH data available.


Related in: MedlinePlus