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Macrophage migration inhibitory factor is overexpressed in pancreatic cancer tissues and impairs insulin secretion function of β-cell.

Tan L, Ye X, Zhou Y, Yu M, Fu Z, Chen R, Zhuang B, Zeng B, Ye H, Gao W, Lin Q, Li Z, Zhou Q, Chen R - J Transl Med (2014)

Bottom Line: MIF expression was significantly increased in pancreatic cancer tissues compared with chronic pancreatitis or normal pancreas specimens.Stable MIF knock-down significantly decreased the diabetogenic effect of PC cells, while MIF knock-in HPDE6 cells demonstrated a strong inhibitory effect on insulin secretion function of islets and HIT-T15 cells.Mean serum level of MIF was significant higher in new-onset diabetes associated PC patients in comparison with other groups.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pancreaticobiliary Surgery, The Second Affiliated Hospital of Sun Yat-sen University (Sun Yat-sen Memorial Hospital), Sun Yat-sen University, 107 Yan-Jiang Xi Road, Guangzhou 510120, China. zqb_sysu@126.com.

ABSTRACT

Background: Understanding the pathogenic mechanism of pancreatic cancer associated diabetes (PCDM) might help yield biomarkers for the early diagnosis of pancreatic cancer (PC) from population with new-onset diabetes. In the current study, we sought to determine the role of macrophage migration inhibitory factor (MIF) in PCDM pathogenesis.

Methods: The protein and mRNA levels of MIF in paraffin-embedded human PC samples, chronic pancreatitis specimens, and normal pancreas were measured by immunohistochemistry and quantitative reverse-transcriptase polymerase chain reaction. We measured serum levels of MIF in PC patients and controls. The biologic impacts of MIF overexpression on insulin secretion function of mice islets and β cells (HIT-T15) were investigated in vitro.

Results: MIF expression was significantly increased in pancreatic cancer tissues compared with chronic pancreatitis or normal pancreas specimens. The insulin secretion function of both islets and HIT-T15 cells was impaired by indirect co-cultured with PC cells or treated with conditioned media from them. Stable MIF knock-down significantly decreased the diabetogenic effect of PC cells, while MIF knock-in HPDE6 cells demonstrated a strong inhibitory effect on insulin secretion function of islets and HIT-T15 cells. MIF impaired βcell function by depressing the Ca⁺ currents, decreasing L-type Ca⁺ channel α1 subunit protein expression level, and enhancing p-Src activity. Mean serum level of MIF was significant higher in new-onset diabetes associated PC patients in comparison with other groups.

Conclusions: MIF is up-regulated in patients with pancreatic cancer and causes dysfunction of insulin secretion in β-cells.

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Related in: MedlinePlus

MIF inhibits glucose-stimulated insulin secretion and contributes to the insulin inhibitory effect of PC cells. (A) Stable shRNA MIF-knockdown (sh-Capan-2, sh-PANC-1) cells and MIF-over expressing cells (MIF-HPDE6) were generated by using lentiviral vectors. Vector controls showed no regulatory effect on MIF expression. (B1&B2) Degree of inhibitory effect on insulin releasing improved with the increasing concentration of MIF from 0 to 100 nmol/L both in PANC-1 (B1) and Capan-2 (B2) cell lines. A most significant drop of insulin secretion level was observed following exposure to MIF at concentrations of 40 nmol/L, reduced to 52% in islets and 46.7% in HIT-T15 cells, respectively, 0 nmol/L rMIF act as control groups. (P <0.01, n = 6). (C1&C2) MIF mediated the inhibitory effect of PC cells on insulin secretion capacity. C1: after indirectly co-cultured with PANC-1 or Capan-2 cells, islet cells showed impaired glucose-stimulated insulin secretion function, shRNA-lentivirus mediated MIF inhibition ameliorated the insulin secretion capacity; the immortalized epithelial cell line HPDE6 decreased insulin secretion a little, while MIF-transgene expressing HPDE6 cells showed a strong inhibitory effect on insulin secretion. C2: after adding supernatants into the culture medium of islet cells, a same effect of MIF was observed. (D1&D2) The same role of MIF was observed in HIT-T15 cells just as it shown in the islet cells. Glc: glucose; *:p<0.01, Student’s t-test. Sh-PANC-1: shRNA MIF-knockdown PANC-1 cell line; Sh-Capan-2: shRNA MIF-knockdown Capan-2 cell line; MIF-HPDE6: MIF-transgene overexpression HPDE6 cell line; pGIZ-PANC-1, pGIZ-Capan-2, and pLOC-HPDE6 are  vector transfected cell lines as controls to exclude the effect of vector on expression of potential genes coding proteins which might regulate insulin releasing.
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Figure 2: MIF inhibits glucose-stimulated insulin secretion and contributes to the insulin inhibitory effect of PC cells. (A) Stable shRNA MIF-knockdown (sh-Capan-2, sh-PANC-1) cells and MIF-over expressing cells (MIF-HPDE6) were generated by using lentiviral vectors. Vector controls showed no regulatory effect on MIF expression. (B1&B2) Degree of inhibitory effect on insulin releasing improved with the increasing concentration of MIF from 0 to 100 nmol/L both in PANC-1 (B1) and Capan-2 (B2) cell lines. A most significant drop of insulin secretion level was observed following exposure to MIF at concentrations of 40 nmol/L, reduced to 52% in islets and 46.7% in HIT-T15 cells, respectively, 0 nmol/L rMIF act as control groups. (P <0.01, n = 6). (C1&C2) MIF mediated the inhibitory effect of PC cells on insulin secretion capacity. C1: after indirectly co-cultured with PANC-1 or Capan-2 cells, islet cells showed impaired glucose-stimulated insulin secretion function, shRNA-lentivirus mediated MIF inhibition ameliorated the insulin secretion capacity; the immortalized epithelial cell line HPDE6 decreased insulin secretion a little, while MIF-transgene expressing HPDE6 cells showed a strong inhibitory effect on insulin secretion. C2: after adding supernatants into the culture medium of islet cells, a same effect of MIF was observed. (D1&D2) The same role of MIF was observed in HIT-T15 cells just as it shown in the islet cells. Glc: glucose; *:p<0.01, Student’s t-test. Sh-PANC-1: shRNA MIF-knockdown PANC-1 cell line; Sh-Capan-2: shRNA MIF-knockdown Capan-2 cell line; MIF-HPDE6: MIF-transgene overexpression HPDE6 cell line; pGIZ-PANC-1, pGIZ-Capan-2, and pLOC-HPDE6 are vector transfected cell lines as controls to exclude the effect of vector on expression of potential genes coding proteins which might regulate insulin releasing.

Mentions: Because MIF was overexpressed in pancreatic cancer cells, we used MIF shRNA to knock down MIF in PANC-1 and Capan-2 cells. Vice versa, a MIF overexpressing immortalized human pancreatic ductal epithelial cell line was generated. The MIF expression was analyzed by western blot and real-time PCR, results showed that stable MIF-over expressing immortalized human pancreatic ductal epithelial cells (HPDE6) and shRNA MIF-knockdown pancreatic cancer cells (PANC-1 and Capan-2) were generated (Figure 2A).


Macrophage migration inhibitory factor is overexpressed in pancreatic cancer tissues and impairs insulin secretion function of β-cell.

Tan L, Ye X, Zhou Y, Yu M, Fu Z, Chen R, Zhuang B, Zeng B, Ye H, Gao W, Lin Q, Li Z, Zhou Q, Chen R - J Transl Med (2014)

MIF inhibits glucose-stimulated insulin secretion and contributes to the insulin inhibitory effect of PC cells. (A) Stable shRNA MIF-knockdown (sh-Capan-2, sh-PANC-1) cells and MIF-over expressing cells (MIF-HPDE6) were generated by using lentiviral vectors. Vector controls showed no regulatory effect on MIF expression. (B1&B2) Degree of inhibitory effect on insulin releasing improved with the increasing concentration of MIF from 0 to 100 nmol/L both in PANC-1 (B1) and Capan-2 (B2) cell lines. A most significant drop of insulin secretion level was observed following exposure to MIF at concentrations of 40 nmol/L, reduced to 52% in islets and 46.7% in HIT-T15 cells, respectively, 0 nmol/L rMIF act as control groups. (P <0.01, n = 6). (C1&C2) MIF mediated the inhibitory effect of PC cells on insulin secretion capacity. C1: after indirectly co-cultured with PANC-1 or Capan-2 cells, islet cells showed impaired glucose-stimulated insulin secretion function, shRNA-lentivirus mediated MIF inhibition ameliorated the insulin secretion capacity; the immortalized epithelial cell line HPDE6 decreased insulin secretion a little, while MIF-transgene expressing HPDE6 cells showed a strong inhibitory effect on insulin secretion. C2: after adding supernatants into the culture medium of islet cells, a same effect of MIF was observed. (D1&D2) The same role of MIF was observed in HIT-T15 cells just as it shown in the islet cells. Glc: glucose; *:p<0.01, Student’s t-test. Sh-PANC-1: shRNA MIF-knockdown PANC-1 cell line; Sh-Capan-2: shRNA MIF-knockdown Capan-2 cell line; MIF-HPDE6: MIF-transgene overexpression HPDE6 cell line; pGIZ-PANC-1, pGIZ-Capan-2, and pLOC-HPDE6 are  vector transfected cell lines as controls to exclude the effect of vector on expression of potential genes coding proteins which might regulate insulin releasing.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4022046&req=5

Figure 2: MIF inhibits glucose-stimulated insulin secretion and contributes to the insulin inhibitory effect of PC cells. (A) Stable shRNA MIF-knockdown (sh-Capan-2, sh-PANC-1) cells and MIF-over expressing cells (MIF-HPDE6) were generated by using lentiviral vectors. Vector controls showed no regulatory effect on MIF expression. (B1&B2) Degree of inhibitory effect on insulin releasing improved with the increasing concentration of MIF from 0 to 100 nmol/L both in PANC-1 (B1) and Capan-2 (B2) cell lines. A most significant drop of insulin secretion level was observed following exposure to MIF at concentrations of 40 nmol/L, reduced to 52% in islets and 46.7% in HIT-T15 cells, respectively, 0 nmol/L rMIF act as control groups. (P <0.01, n = 6). (C1&C2) MIF mediated the inhibitory effect of PC cells on insulin secretion capacity. C1: after indirectly co-cultured with PANC-1 or Capan-2 cells, islet cells showed impaired glucose-stimulated insulin secretion function, shRNA-lentivirus mediated MIF inhibition ameliorated the insulin secretion capacity; the immortalized epithelial cell line HPDE6 decreased insulin secretion a little, while MIF-transgene expressing HPDE6 cells showed a strong inhibitory effect on insulin secretion. C2: after adding supernatants into the culture medium of islet cells, a same effect of MIF was observed. (D1&D2) The same role of MIF was observed in HIT-T15 cells just as it shown in the islet cells. Glc: glucose; *:p<0.01, Student’s t-test. Sh-PANC-1: shRNA MIF-knockdown PANC-1 cell line; Sh-Capan-2: shRNA MIF-knockdown Capan-2 cell line; MIF-HPDE6: MIF-transgene overexpression HPDE6 cell line; pGIZ-PANC-1, pGIZ-Capan-2, and pLOC-HPDE6 are vector transfected cell lines as controls to exclude the effect of vector on expression of potential genes coding proteins which might regulate insulin releasing.
Mentions: Because MIF was overexpressed in pancreatic cancer cells, we used MIF shRNA to knock down MIF in PANC-1 and Capan-2 cells. Vice versa, a MIF overexpressing immortalized human pancreatic ductal epithelial cell line was generated. The MIF expression was analyzed by western blot and real-time PCR, results showed that stable MIF-over expressing immortalized human pancreatic ductal epithelial cells (HPDE6) and shRNA MIF-knockdown pancreatic cancer cells (PANC-1 and Capan-2) were generated (Figure 2A).

Bottom Line: MIF expression was significantly increased in pancreatic cancer tissues compared with chronic pancreatitis or normal pancreas specimens.Stable MIF knock-down significantly decreased the diabetogenic effect of PC cells, while MIF knock-in HPDE6 cells demonstrated a strong inhibitory effect on insulin secretion function of islets and HIT-T15 cells.Mean serum level of MIF was significant higher in new-onset diabetes associated PC patients in comparison with other groups.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pancreaticobiliary Surgery, The Second Affiliated Hospital of Sun Yat-sen University (Sun Yat-sen Memorial Hospital), Sun Yat-sen University, 107 Yan-Jiang Xi Road, Guangzhou 510120, China. zqb_sysu@126.com.

ABSTRACT

Background: Understanding the pathogenic mechanism of pancreatic cancer associated diabetes (PCDM) might help yield biomarkers for the early diagnosis of pancreatic cancer (PC) from population with new-onset diabetes. In the current study, we sought to determine the role of macrophage migration inhibitory factor (MIF) in PCDM pathogenesis.

Methods: The protein and mRNA levels of MIF in paraffin-embedded human PC samples, chronic pancreatitis specimens, and normal pancreas were measured by immunohistochemistry and quantitative reverse-transcriptase polymerase chain reaction. We measured serum levels of MIF in PC patients and controls. The biologic impacts of MIF overexpression on insulin secretion function of mice islets and β cells (HIT-T15) were investigated in vitro.

Results: MIF expression was significantly increased in pancreatic cancer tissues compared with chronic pancreatitis or normal pancreas specimens. The insulin secretion function of both islets and HIT-T15 cells was impaired by indirect co-cultured with PC cells or treated with conditioned media from them. Stable MIF knock-down significantly decreased the diabetogenic effect of PC cells, while MIF knock-in HPDE6 cells demonstrated a strong inhibitory effect on insulin secretion function of islets and HIT-T15 cells. MIF impaired βcell function by depressing the Ca⁺ currents, decreasing L-type Ca⁺ channel α1 subunit protein expression level, and enhancing p-Src activity. Mean serum level of MIF was significant higher in new-onset diabetes associated PC patients in comparison with other groups.

Conclusions: MIF is up-regulated in patients with pancreatic cancer and causes dysfunction of insulin secretion in β-cells.

Show MeSH
Related in: MedlinePlus