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sCLU regulates cisplatin chemosensitivity of lung cancer cells in vivo.

Ma G, Cai H, Gao L, Wang M, Wang H - World J Surg Oncol (2015)

Bottom Line: The effect of sCLU silencing or overexpression on pAkt and pERK1/2 expression and chemosensitivity in vivo was detected by Western blot assay.The results showed sCLU silencing increased the chemosensitivity of A549(DDP) cells to DDP in vivo via downregulation of pAkt and pERK1/2 expression.And sCLU overexpression decreased the chemosensitivity of A549 cells to DDP in vivo via upregulation of pAkt and pERK1/2 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Lab, People's Hospital of Laiwu, Laiwu, China. maguoliang7782@sina.com.

ABSTRACT

Background: In a previous analysis using a lung cancer cell line model, we have found that therapies directed against secreted clusterin (sCLU) and its downstream signaling targets pAkt and pERK1/2 may have the potential to enhance the efficacy of cisplatin (DDP)-based chemotherapy in vitro. Here, we investigated the therapies directed against sCLU on the DDP-based chemotherapy in vivo and explored the mechanism.

Methods: Using lung cancer cell lines, A549 cells and DDP-resistant A549 cells (A549(DDP)), we determined the effect of sCLU silencing using short interfering double-stranded RNA (siRNA) on chemosensitivity in immunocompromised mice bearing A549(DDP) tumors. We then determined the effect of sCLU overexpression via stable sCLU transfection on chemosensitivity in immunocompromised mice bearing A549 tumors. The effect of sCLU silencing or overexpression on pAkt and pERK1/2 expression and chemosensitivity in vivo was detected by Western blot assay.

Results: The results showed sCLU silencing increased the chemosensitivity of A549(DDP) cells to DDP in vivo via downregulation of pAkt and pERK1/2 expression. And sCLU overexpression decreased the chemosensitivity of A549 cells to DDP in vivo via upregulation of pAkt and pERK1/2 expression.

Conclusions: We therefore concluded that the DDP-induced sCLU activation, which involved induction of pAkt and pERK1/2 activation that confer DDP resistance in immunocompromised mice and alteration of this balance, allows sensitization to the antitumor activity of cisplatin chemotherapy.

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Clusterin overexpression decreased chemotherapeutic sensitivity and inhibited apoptosis of DDP sensitive A549 cells. (A) Cells transfected with pCDNA3.1 /sCLU or scramble pCDNA3.1 were injected subcutaneously into the right flank of nude mice. When tumors reached −100 mm3 in volume, the mice received daily 200 μl i.p. injections of DDP (4 mg/kg body/wt.,i.p). DDP was administered i.v. once every 3 days. The treatments lasted for 15 days during which the volume of tumors was recorded,*P < 0.05(vs pCDNA3.1 /sCLU + DDP) (Student’s t test). (B) Tumor sections were stained with TUNEL agent to visualize apoptotic cells. *P < 0.05 (Student’s t test).
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Fig1: Clusterin overexpression decreased chemotherapeutic sensitivity and inhibited apoptosis of DDP sensitive A549 cells. (A) Cells transfected with pCDNA3.1 /sCLU or scramble pCDNA3.1 were injected subcutaneously into the right flank of nude mice. When tumors reached −100 mm3 in volume, the mice received daily 200 μl i.p. injections of DDP (4 mg/kg body/wt.,i.p). DDP was administered i.v. once every 3 days. The treatments lasted for 15 days during which the volume of tumors was recorded,*P < 0.05(vs pCDNA3.1 /sCLU + DDP) (Student’s t test). (B) Tumor sections were stained with TUNEL agent to visualize apoptotic cells. *P < 0.05 (Student’s t test).

Mentions: Based on the in vitro experiment of clusterin in the cisplatin resistance of lung cancers [11], we further examined if clusterin expression affects the cisplatin sensitivity in vivo; A549, A549/sCLU, and A549/pcDNA3.1 cells were injected subcutaneously into the right flank of nude mice. Cisplatin could significantly inhibit the tumor growth in mice injected with A549/pCDNA3.1 scramble cells and A549 cells compared to the mice injected with A549/sCLU cells (Figure 1A). As shown in Figure 1A, the A549 tumors of mice treated with DDP only reached 530 ± 18.6 mm3 in volume 36 days after treatment, which was significantly smaller compared to A549/sCLU cells (1184.4 ± 102.6 mm3) in volume 36 days after DDP treatment (P < 0.05). Clusterin overexpression alone showed no significant growth inhibition compared to the control group.Figure 1


sCLU regulates cisplatin chemosensitivity of lung cancer cells in vivo.

Ma G, Cai H, Gao L, Wang M, Wang H - World J Surg Oncol (2015)

Clusterin overexpression decreased chemotherapeutic sensitivity and inhibited apoptosis of DDP sensitive A549 cells. (A) Cells transfected with pCDNA3.1 /sCLU or scramble pCDNA3.1 were injected subcutaneously into the right flank of nude mice. When tumors reached −100 mm3 in volume, the mice received daily 200 μl i.p. injections of DDP (4 mg/kg body/wt.,i.p). DDP was administered i.v. once every 3 days. The treatments lasted for 15 days during which the volume of tumors was recorded,*P < 0.05(vs pCDNA3.1 /sCLU + DDP) (Student’s t test). (B) Tumor sections were stained with TUNEL agent to visualize apoptotic cells. *P < 0.05 (Student’s t test).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4358856&req=5

Fig1: Clusterin overexpression decreased chemotherapeutic sensitivity and inhibited apoptosis of DDP sensitive A549 cells. (A) Cells transfected with pCDNA3.1 /sCLU or scramble pCDNA3.1 were injected subcutaneously into the right flank of nude mice. When tumors reached −100 mm3 in volume, the mice received daily 200 μl i.p. injections of DDP (4 mg/kg body/wt.,i.p). DDP was administered i.v. once every 3 days. The treatments lasted for 15 days during which the volume of tumors was recorded,*P < 0.05(vs pCDNA3.1 /sCLU + DDP) (Student’s t test). (B) Tumor sections were stained with TUNEL agent to visualize apoptotic cells. *P < 0.05 (Student’s t test).
Mentions: Based on the in vitro experiment of clusterin in the cisplatin resistance of lung cancers [11], we further examined if clusterin expression affects the cisplatin sensitivity in vivo; A549, A549/sCLU, and A549/pcDNA3.1 cells were injected subcutaneously into the right flank of nude mice. Cisplatin could significantly inhibit the tumor growth in mice injected with A549/pCDNA3.1 scramble cells and A549 cells compared to the mice injected with A549/sCLU cells (Figure 1A). As shown in Figure 1A, the A549 tumors of mice treated with DDP only reached 530 ± 18.6 mm3 in volume 36 days after treatment, which was significantly smaller compared to A549/sCLU cells (1184.4 ± 102.6 mm3) in volume 36 days after DDP treatment (P < 0.05). Clusterin overexpression alone showed no significant growth inhibition compared to the control group.Figure 1

Bottom Line: The effect of sCLU silencing or overexpression on pAkt and pERK1/2 expression and chemosensitivity in vivo was detected by Western blot assay.The results showed sCLU silencing increased the chemosensitivity of A549(DDP) cells to DDP in vivo via downregulation of pAkt and pERK1/2 expression.And sCLU overexpression decreased the chemosensitivity of A549 cells to DDP in vivo via upregulation of pAkt and pERK1/2 expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Clinical Lab, People's Hospital of Laiwu, Laiwu, China. maguoliang7782@sina.com.

ABSTRACT

Background: In a previous analysis using a lung cancer cell line model, we have found that therapies directed against secreted clusterin (sCLU) and its downstream signaling targets pAkt and pERK1/2 may have the potential to enhance the efficacy of cisplatin (DDP)-based chemotherapy in vitro. Here, we investigated the therapies directed against sCLU on the DDP-based chemotherapy in vivo and explored the mechanism.

Methods: Using lung cancer cell lines, A549 cells and DDP-resistant A549 cells (A549(DDP)), we determined the effect of sCLU silencing using short interfering double-stranded RNA (siRNA) on chemosensitivity in immunocompromised mice bearing A549(DDP) tumors. We then determined the effect of sCLU overexpression via stable sCLU transfection on chemosensitivity in immunocompromised mice bearing A549 tumors. The effect of sCLU silencing or overexpression on pAkt and pERK1/2 expression and chemosensitivity in vivo was detected by Western blot assay.

Results: The results showed sCLU silencing increased the chemosensitivity of A549(DDP) cells to DDP in vivo via downregulation of pAkt and pERK1/2 expression. And sCLU overexpression decreased the chemosensitivity of A549 cells to DDP in vivo via upregulation of pAkt and pERK1/2 expression.

Conclusions: We therefore concluded that the DDP-induced sCLU activation, which involved induction of pAkt and pERK1/2 activation that confer DDP resistance in immunocompromised mice and alteration of this balance, allows sensitization to the antitumor activity of cisplatin chemotherapy.

Show MeSH
Related in: MedlinePlus