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Extraction of Peptidoglycan from L. paracasei subp. Paracasei X12 and Its Preliminary Mechanisms of Inducing Immunogenic Cell Death in HT-29 Cells.

Tian PJ, Li BL, Shan YJ, Zhang JN, Chen JY, Yu M, Zhang LW - Int J Mol Sci (2015)

Bottom Line: X12-PG could induce the production of apoptotic bodies observed by transmission electron microscopy (TEM).X12-PG could significantly induced the translocation of calreticulin (CRT) and the release of high mobility group box 1 protein (HMGB1), the two notable hallmarks of immunogenic cell death (ICD), with the endoplastic reticulum (ER) damaged and subsequently intracellular [Ca(2+)] elevated.Our findings implied that X12-PG could induce the ICD of HT-29 cells through targeting at the ER.

View Article: PubMed Central - PubMed

Affiliation: School of Food Science and Engineering, Harbin Institute of Technology, No. 73 Huanghe Road, Harbin 150000, China. tianpei0202@gmail.com.

ABSTRACT
L. paracasei subp. paracasei X12 was previously isolated from a Chinese traditional fermented cheese with anticancer activities and probiotic potential. Herein, the integral peptidoglycan (X12-PG) was extracted by a modified trichloroacetic acid (TCA) method. X12-PG contained the four representative amino acids Asp, Glu, Ala and Lys, and displayed the similar lysozyme sensitivity, UV-visible scanning spectrum and molecular weight as the peptidoglycan standard. X12-PG could induce the production of apoptotic bodies observed by transmission electron microscopy (TEM). X12-PG could significantly induced the translocation of calreticulin (CRT) and the release of high mobility group box 1 protein (HMGB1), the two notable hallmarks of immunogenic cell death (ICD), with the endoplastic reticulum (ER) damaged and subsequently intracellular [Ca(2+)] elevated. Our findings implied that X12-PG could induce the ICD of HT-29 cells through targeting at the ER. The present results may enlighten the prospect of probiotics in the prevention of colon cancer.

No MeSH data available.


Related in: MedlinePlus

Apoptosis of HT-29 cells after treatment of X12-PG. (A–D) were respectively treated with 0, 400, 800, 1600 μg/mL X12-PG for 48 h. HT-29 cells were observed by TEM (×6000). The arrows indicate the apoptotic bodies.
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ijms-16-20033-f003: Apoptosis of HT-29 cells after treatment of X12-PG. (A–D) were respectively treated with 0, 400, 800, 1600 μg/mL X12-PG for 48 h. HT-29 cells were observed by TEM (×6000). The arrows indicate the apoptotic bodies.

Mentions: Morphological studies of X12-PG-treated and untreated HT-29 cells were carried out by TEM. In untreated cells, a large number of villi-like projections on cell surface were distributed uniformly. In addition, nuclear chromatin and intact organelles such as endoplasmic reticulum, mitochondria and ribosomes, were clearly observed (Figure 3A). In X12-PG-treated cells, however, villous structures were gradually diminished, meanwhile accompanied by mitochondria degradation, increased cytoplasmic vacuoles and formation of apoptotic bodies (Figure 3B,C, arrows showed apoptotic body). This alteration was enhanced when the treatment concentration increased, especially at 1600 μg/mL, villous structures almost completely disappeared. Except for mature apoptotic bodies, cell shrinkage and increased nuclear/cytoplasmic ratio were also observed (Figure 3D).


Extraction of Peptidoglycan from L. paracasei subp. Paracasei X12 and Its Preliminary Mechanisms of Inducing Immunogenic Cell Death in HT-29 Cells.

Tian PJ, Li BL, Shan YJ, Zhang JN, Chen JY, Yu M, Zhang LW - Int J Mol Sci (2015)

Apoptosis of HT-29 cells after treatment of X12-PG. (A–D) were respectively treated with 0, 400, 800, 1600 μg/mL X12-PG for 48 h. HT-29 cells were observed by TEM (×6000). The arrows indicate the apoptotic bodies.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4581339&req=5

ijms-16-20033-f003: Apoptosis of HT-29 cells after treatment of X12-PG. (A–D) were respectively treated with 0, 400, 800, 1600 μg/mL X12-PG for 48 h. HT-29 cells were observed by TEM (×6000). The arrows indicate the apoptotic bodies.
Mentions: Morphological studies of X12-PG-treated and untreated HT-29 cells were carried out by TEM. In untreated cells, a large number of villi-like projections on cell surface were distributed uniformly. In addition, nuclear chromatin and intact organelles such as endoplasmic reticulum, mitochondria and ribosomes, were clearly observed (Figure 3A). In X12-PG-treated cells, however, villous structures were gradually diminished, meanwhile accompanied by mitochondria degradation, increased cytoplasmic vacuoles and formation of apoptotic bodies (Figure 3B,C, arrows showed apoptotic body). This alteration was enhanced when the treatment concentration increased, especially at 1600 μg/mL, villous structures almost completely disappeared. Except for mature apoptotic bodies, cell shrinkage and increased nuclear/cytoplasmic ratio were also observed (Figure 3D).

Bottom Line: X12-PG could induce the production of apoptotic bodies observed by transmission electron microscopy (TEM).X12-PG could significantly induced the translocation of calreticulin (CRT) and the release of high mobility group box 1 protein (HMGB1), the two notable hallmarks of immunogenic cell death (ICD), with the endoplastic reticulum (ER) damaged and subsequently intracellular [Ca(2+)] elevated.Our findings implied that X12-PG could induce the ICD of HT-29 cells through targeting at the ER.

View Article: PubMed Central - PubMed

Affiliation: School of Food Science and Engineering, Harbin Institute of Technology, No. 73 Huanghe Road, Harbin 150000, China. tianpei0202@gmail.com.

ABSTRACT
L. paracasei subp. paracasei X12 was previously isolated from a Chinese traditional fermented cheese with anticancer activities and probiotic potential. Herein, the integral peptidoglycan (X12-PG) was extracted by a modified trichloroacetic acid (TCA) method. X12-PG contained the four representative amino acids Asp, Glu, Ala and Lys, and displayed the similar lysozyme sensitivity, UV-visible scanning spectrum and molecular weight as the peptidoglycan standard. X12-PG could induce the production of apoptotic bodies observed by transmission electron microscopy (TEM). X12-PG could significantly induced the translocation of calreticulin (CRT) and the release of high mobility group box 1 protein (HMGB1), the two notable hallmarks of immunogenic cell death (ICD), with the endoplastic reticulum (ER) damaged and subsequently intracellular [Ca(2+)] elevated. Our findings implied that X12-PG could induce the ICD of HT-29 cells through targeting at the ER. The present results may enlighten the prospect of probiotics in the prevention of colon cancer.

No MeSH data available.


Related in: MedlinePlus