caprin2 sirna  (Santa Cruz Biotechnology)

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: Effects of CAPRIN2 on the ferroptosis and survival of ECM-detached NPC cells and the migration and invasion of NPC cells. (A) Viability assay of ECM-detached NPC cell lines treated with erastin (5 μM) and/or ferrostatin-1 (1 μM) for 24 h (B) Viability assay of the indicated stable 5-8F or C666-1 cell lines cultured under ECM detachment conditions for 72 h For (A, B) , the experiments were repeated three times, and the data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. (C, D) Migration and invasion assays of the indicated stable 5-8F (C) and C666-1 (D) cell lines. These assays were conducted in triplicate. Representative images are displayed. The data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Migration, Viability Assay, Cell Culture

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: CAPRIN2 promotes the ferroptosis resistance, survival, migration and invasion of NPC cells through HMGCR. (A) Overexpression of HMGCR partially reverses the effects of CAPRIN2 on the ferroptosis of ECM-detached 5-8F (left panel) and C666-1 (right panel) cells. The NPC cell lines were treated with erastin (5 μM) for 24 h (B, C) MDA assay (B) and GSH assay (C) results of erastin-treated NPC stable cell lines as indicated. (D) Ectopic expression of HMGCR partially rescues the effects of CAPRIN2 knockdown on ECM-detached NPC cell survival. For (A–C) and (D) , the experiments were conducted in triplicate, and the data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01. (E, F) Stable overexpression of HMGCR partially reverses the effects of CAPRIN2 knockdown on 5-8F (E) and C666-1 (F) cell migration and invasion. Representative images of three independent experiments are shown. The data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Migration, Over Expression, Multiple Displacement Amplification, GSH Assay, Stable Transfection, Expressing, Knockdown

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: Regulation of lung colonization capacity via the CAPRIN2/HMGCR axis in NPC cells. (A, B) The inhibitory effect of erastin on the lung metastasis of NPC cells was enhanced by knockdown of the CAPRIN2/HMGCR axis. (C, D) CAPRIN2 promotes the lung colonization of NPC cells through HMGCR. For (A, C) , representative images of lungs and HE staining are shown. The location of lung metastatic nodules is indicated by the arrow. For (B, D) , the number of lung metastases (left panel) and the weight of the lungs (right panel) are given. * p < 0.05, ** p < 0.01.

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Knockdown, Staining

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: LINC00941 acts as an upstream molecule to regulate the biological functions of CAPRIN2. (A) LINC00941 downregulation promoted the ferroptosis of ECM-detached NPC cells, which was partially rescued by CAPRIN2 overexpression. The NPC cells were treated with erastin (5 μM) for 24 h (B) Knockdown of LINC00941 decreased the survival of ECM-detached NPC cells, which could be partially reversed by CAPRIN2 overexpression. For (A) and (B) , the assays were conducted in triplicate, and the data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01. (C, D) LINC00941 knockdown inhibited the migration and invasion of 5-8F (C) and C666-1 (D) cells, and overexpression of CAPRIN2 partially reversed this effect. Representative images are shown. The data are provided as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Over Expression, Knockdown, Migration

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: CAPRIN2 is overactivated in NPC tissues and is associated with a poor prognosis in patients. (A) The expression level of CAPRIN2 in 20 NPC tissues and 5 nasopharyngeal tissues. (B) The correlation between LINC00941/CAPRIN2, CAPRIN2/HMGCR or LINC00941/HMGCR in 20 NPC tissues. For (A) and (B) , the expression levels of CAPRIN2, HMGCR and LINC00941 were determined by qRT-PCR. The levels were normalized to those of β-actin and shown as the mean ± SEM. * p < 0.05. (C) Representative immunohistochemical images of normal nasopharyngeal tissues (left panel) and NPC tissues (middle panel). The boxes represent the magnified region. The representative image of negative stained control (right panel, top) shows the negative staining result of NPC tissues incubated with antibody-free serum. The representative image of positive stained control (right panel, bottom) shows the positive staining result of CAPRIN2 in NPC tissues incubated with the primary antibody of CAPRIN2. (D) Kaplan-Meier survival analysis of the association between CAPRIN2 expression and the PFS or OS of NPC patients ( log-rank test ).

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Staining, Control, Negative Staining, Incubation

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: Correlations between CAPRIN2 expression and clinicopathological characteristics.

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: Univariate and multivariate analysis for OS.

Article Snippet: The siRNAs applied in the study were all products of Santa Cruz (Shanghai, China) and are listed as follows: CAPRIN2 siRNA, HMGCR siRNA and negative control siRNA.

Techniques: Expressing

Journal: Frontiers in oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR.

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: FIGURE 1 Effects of CAPRIN2 on the ferroptosis and survival of ECM-detached NPC cells and the migration and invasion of NPC cells. (A) Viability assay of ECM-detached NPC cell lines treated with erastin (5 mM) and/or ferrostatin-1 (1 mM) for 24 h (B) Viability assay of the indicated stable 5-8F or C666-1 cell lines cultured under ECM detachment conditions for 72 h For (A, B), the experiments were repeated three times, and the data are shown as the mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001. (C, D) Migration and invasion assays of the indicated stable 5-8F (C) and C666-1 (D) cell lines. These assays were conducted in triplicate. Representative images are displayed. The data are presented as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Stable cell line construction To construct stable cell lines with knockdown of CAPRIN2 or HMGCR, lentiviruses carrying CAPRIN2 shRNA, HMGCR shRNA or scramble shRNA were purchased from Santa Cruz (CA, USA) and used to infect the indicated NPC cell lines for 48 h. The sequence of human LINC00941 shRNA was 5′- GAGACAGTTGATAGCCAAA -3′ (36), and the constructs were cloned into pHBLV-U6-MCS-PGK-PURO, named pHBLV-shLINC00941. pHBLV-shLINC00941 was transfected into 293T cells along with the corresponding packaging vector PMD2.G and pSPAX2.

Techniques: Migration, Viability Assay, Cell Culture

Journal: Frontiers in oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR.

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: FIGURE 2 CAPRIN2 promotes the ferroptosis resistance, survival, migration and invasion of NPC cells through HMGCR. (A) Overexpression of HMGCR partially reverses the effects of CAPRIN2 on the ferroptosis of ECM-detached 5-8F (left panel) and C666-1 (right panel) cells. The NPC cell lines were treated with erastin (5 mM) for 24 h (B, C) MDA assay (B) and GSH assay (C) results of erastin-treated NPC stable cell lines as indicated. (D) Ectopic expression of HMGCR partially rescues the effects of CAPRIN2 knockdown on ECM-detached NPC cell survival. For (A–C) and (D), the experiments were conducted in triplicate, and the data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01. (E, F) Stable overexpression of HMGCR partially reverses the effects of CAPRIN2 knockdown on 5-8F (E) and C666-1 (F) cell migration and invasion. Representative images of three independent experiments are shown. The data are expressed as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Stable cell line construction To construct stable cell lines with knockdown of CAPRIN2 or HMGCR, lentiviruses carrying CAPRIN2 shRNA, HMGCR shRNA or scramble shRNA were purchased from Santa Cruz (CA, USA) and used to infect the indicated NPC cell lines for 48 h. The sequence of human LINC00941 shRNA was 5′- GAGACAGTTGATAGCCAAA -3′ (36), and the constructs were cloned into pHBLV-U6-MCS-PGK-PURO, named pHBLV-shLINC00941. pHBLV-shLINC00941 was transfected into 293T cells along with the corresponding packaging vector PMD2.G and pSPAX2.

Techniques: Migration, Over Expression, Multiple Displacement Amplification, GSH Assay, Stable Transfection, Expressing, Knockdown

Journal: Frontiers in oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR.

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: FIGURE 3 Regulation of lung colonization capacity via the CAPRIN2/HMGCR axis in NPC cells. (A, B) The inhibitory effect of erastin on the lung metastasis of NPC cells was enhanced by knockdown of the CAPRIN2/HMGCR axis. (C, D) CAPRIN2 promotes the lung colonization of NPC cells through HMGCR. For (A, C), representative images of lungs and HE staining are shown. The location of lung metastatic nodules is indicated by the arrow. For (B, D), the number of lung metastases (left panel) and the weight of the lungs (right panel) are given. * p < 0.05, ** p < 0.01.

Article Snippet: Stable cell line construction To construct stable cell lines with knockdown of CAPRIN2 or HMGCR, lentiviruses carrying CAPRIN2 shRNA, HMGCR shRNA or scramble shRNA were purchased from Santa Cruz (CA, USA) and used to infect the indicated NPC cell lines for 48 h. The sequence of human LINC00941 shRNA was 5′- GAGACAGTTGATAGCCAAA -3′ (36), and the constructs were cloned into pHBLV-U6-MCS-PGK-PURO, named pHBLV-shLINC00941. pHBLV-shLINC00941 was transfected into 293T cells along with the corresponding packaging vector PMD2.G and pSPAX2.

Techniques: Knockdown, Staining

Journal: Frontiers in oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR.

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: FIGURE 4 LINC00941 acts as an upstream molecule to regulate the biological functions of CAPRIN2. (A) LINC00941 downregulation promoted the ferroptosis of ECM-detached NPC cells, which was partially rescued by CAPRIN2 overexpression. The NPC cells were treated with erastin (5 mM) for 24 h (B) Knockdown of LINC00941 decreased the survival of ECM-detached NPC cells, which could be partially reversed by CAPRIN2 overexpression. For (A) and (B), the assays were conducted in triplicate, and the data are presented as the mean ± SEM. * p < 0.05, ** p < 0.01. (C, D) LINC00941 knockdown inhibited the migration and invasion of 5-8F (C) and C666-1 (D) cells, and overexpression of CAPRIN2 partially reversed this effect. Representative images are shown. The data are provided as the mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Stable cell line construction To construct stable cell lines with knockdown of CAPRIN2 or HMGCR, lentiviruses carrying CAPRIN2 shRNA, HMGCR shRNA or scramble shRNA were purchased from Santa Cruz (CA, USA) and used to infect the indicated NPC cell lines for 48 h. The sequence of human LINC00941 shRNA was 5′- GAGACAGTTGATAGCCAAA -3′ (36), and the constructs were cloned into pHBLV-U6-MCS-PGK-PURO, named pHBLV-shLINC00941. pHBLV-shLINC00941 was transfected into 293T cells along with the corresponding packaging vector PMD2.G and pSPAX2.

Techniques: Over Expression, Knockdown, Migration

Journal: Frontiers in oncology

Article Title: CAPRIN2 upregulation by LINC00941 promotes nasopharyngeal carcinoma ferroptosis resistance and metastatic colonization through HMGCR.

doi: 10.3389/fonc.2022.931749

Figure Lengend Snippet: FIGURE 5 CAPRIN2 is overactivated in NPC tissues and is associated with a poor prognosis in patients. (A) The expression level of CAPRIN2 in 20 NPC tissues and 5 nasopharyngeal tissues. (B) The correlation between LINC00941/CAPRIN2, CAPRIN2/HMGCR or LINC00941/HMGCR in 20 NPC tissues. For (A) and (B), the expression levels of CAPRIN2, HMGCR and LINC00941 were determined by qRT-PCR. The levels were normalized to those of b-actin and shown as the mean ± SEM. * p < 0.05. (C) Representative immunohistochemical images of normal nasopharyngeal tissues (left panel) and NPC tissues (middle panel). The boxes represent the magnified region. The representative image of negative stained control (right panel, top) shows the negative staining result of NPC tissues incubated with antibody-free serum. The representative image of positive stained control (right panel, bottom) shows the positive staining result of CAPRIN2 in NPC tissues incubated with the primary antibody of CAPRIN2. (D) Kaplan-Meier survival analysis of the association between CAPRIN2 expression and the PFS or OS of NPC patients (log-rank test).

Article Snippet: Stable cell line construction To construct stable cell lines with knockdown of CAPRIN2 or HMGCR, lentiviruses carrying CAPRIN2 shRNA, HMGCR shRNA or scramble shRNA were purchased from Santa Cruz (CA, USA) and used to infect the indicated NPC cell lines for 48 h. The sequence of human LINC00941 shRNA was 5′- GAGACAGTTGATAGCCAAA -3′ (36), and the constructs were cloned into pHBLV-U6-MCS-PGK-PURO, named pHBLV-shLINC00941. pHBLV-shLINC00941 was transfected into 293T cells along with the corresponding packaging vector PMD2.G and pSPAX2.

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Staining, Control, Negative Staining, Incubation