Relationship of EGFR-TKI Targeted Therapy and Pyroptosis in Non-small Cell Lung Cancer / 肿瘤防治研究

Objective To explore the relationship between pyroptosis and treatment in non-small cell lung cancer patients treated with tyrosine kinase inhibitors targeted therapy. Methods Stable transfection strains with common EGFR mutations found in clinical practice were constructed through lentiviral transfection. LDH and Western blot experiments were conducted to determine the degree and mechanism of pyroptosis after osimertinib treatment. Animal experiments verified the effect of pyroptosis on treatment efficacy. ELISA was used to explore the potential connection between pyroptosis and tumor immunotherapy. Results After osimertinib treatment on stable lines, the EGFR-L858R mutation had obvious pyroptosis at the morphology and protein levels. Western blot experiment confirmed that pyroptosis was mediated by GSDME (P < 0.0001). Experiments through the overexpression of GSDME and corresponding animal studies discovered that the degree of pyroptosis affected the treatment outcome. Blood analysis revealed that the level of IL-1β secreted by EGFR-L858R and EGFR-L858R-GSDME-OE mice after treatment was higher than that of the control group (P < 0.0001), and it may regulate tumor immunity to a certain extent. Conclusion Osimertinib can induce pyroptosis in EGFR-L858R mutant strains mediated by GSDME, and the level of pyroptosis in cell lines is positively correlated with therapeutic effect to a certain extent.

NDFIP1 limits cellular TAZ accumulation via exosomal sorting to inhibit NSCLC proliferation

NDFIP1 has been previously reported as a tumor suppressor in multiple solid tumors, but the function of NDFIP1 in NSCLC and the underlying mechanism are still unknown. Besides, the WW domain containing proteins can be recognized by NDFIP1, resulted in the loading of the target proteins into exosomes. However, whether WW domain-containing transcription regulator 1 (WWTR1, also known as TAZ) can be packaged into exosomes by NDFIP1 and if so, whether the release of this oncogenic protein via exosomes has an effect on tumor development has not been investigated to any extent. Here, we first found that NDFIP1 was low expressed in NSCLC samples and cell lines, which is associated with shorter OS. Then, we confirmed the interaction between TAZ and NDFIP1, and the existence of TAZ in exosomes, which requires NDFIP1. Critically, knockout of NDFIP1 led to TAZ accumulation with no change in its mRNA level and degradation rate. And the cellular TAZ level could be altered by exosome secretion. Furthermore, NDFIP1 inhibited proliferation in vitro and in vivo, and silencing TAZ eliminated the increase of proliferation caused by NDFIP1 knockout. Moreover, TAZ was negatively correlated with NDFIP1 in subcutaneous xenograft model and clinical samples, and the serum exosomal TAZ level was lower in NSCLC patients. In summary, our data uncover a new tumor suppressor, NDFIP1 in NSCLC, and a new exosome-related regulatory mechanism of TAZ.

Expression of acetyl coenzyme A synthetase 2 in colorectal cancer and its biological role / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of acetyl coenzyme A synthetase short-chain family member 2 (ACSS2) in patients with colorectal cancer (CRC) and its biological role.</p><p><b>METHODS</b>A total of 74 CRC tissue samples and 40 normal colorectal tissues were tested by immunohistochemical staining to detect the expression of ACSS2 (cell staining intensity score: 0 points: without staining, 1 points: weak staining, 2 points: intensity staining, 3 point: strong staining; the percentage of positive cells in tumor or negative score: 0 points: negative, 1 point: <25% positive cells, 2 points: 25%-50% positive cells, 3 points: 50%-75% positive cells, 4 points: >75% positive cells. The product of above two scores was the final score.). Association of ACSS2 expression with clinicopathological characteristics was analyzed. Small interfering RNA (siRNA, including A and B group) was used to knock down the expression of ACSS2 in colorectal cell lines (Lovo, HCT116) and their proliferation, migration and epithelial-mesenchymal transition (E-cadherin and Snail as markers) after knocking down were observed.</p><p><b>RESULTS</b>The expression of ACSS2 was significant higher in CRC tissue than that in normal colorectal tissue (tumor average score 6.284, normal tissue average score 3.625, P<0.01) and the percentage of positive cell was higher than that in normal tissue (tumor 69.9%, normal tissue 45.1%, P=0.000). The use of ACSS2 siRNA successfully knocked down the expression of ACSS2 in Lovo and HCT116 cells. A mild suppression of cell proliferation was observed 5 days after planked (A450 value: Lovo-NC 1.758±0.041, Lovo-ACSS2-siA 1.485±10.026, Lovo-ACSS2-siB 1.371±0.049; HCT116-NC 2.609±0.038, HCT116-ACSS2-siA 2.260±0.042, HCT116-ACSS2-siB 2.295±0.029). While a remarkable ability decline of cell migration was found (Lovo-NC 225±5/field, Lovo-ACSS2-siA 40±5/field, Lovo-ACSS2-siB 79±3/field; HCT116-NC 198±7/field, HCT116-ACSS2-siA 96±7/field, HCT116-ACSS2-siB 77±9/field, P<0.05). Real-time quantitative PCR detection showed that in Lovo cells, expression of E-cadherin up-regulated and expression of Snail down-regulated, while in HCT116 cells, E-cadherin up-regulated slightly [E-cadherin: Lovo NC 1.000±0.211, Lovo-siA 3.403±0.207, Lovo-siB 2.658±0.420 (P<0.05); HCT116 NC 1.000±0.121, HCT116-siA 1.349±0.197, HCT116-siB 1.528±0.147(P>0.05); Snail: Lovo NC 1.000±0.085, Lovo-siA 0.468±0.030, Lovo-siB 0.499±0.088 (P<0.05); HCT116 NC 1.000±0.118, HCT116-siA 0.265±0.020, HCT116-siB 0.194±0.017 (P<0.05)].</p><p><b>CONCLUSION</b>CRC tissues have high expression of ACSS2, which may be associated with cell migration and epithelial-mesenchymal transition.</p>

Functional electrical stimulation for improving motor function in elderly patients with cerebral apoplexy hemiplegia / 中华物理医学与康复杂志

Objective Electric stimulation functional observation statistics (FES) were collected for elderly patients with cerebral apoplexy in search of any curative effect. Methods A total of 40 patients with cerebral apoplexy were divided randomly into a FES group of 20 cases and a control group of 20 cases. Both groups received the same routine rehabilitation training and basic drugs. The FES group was also treated using FES therapeutic apparatus for 30 min daily over 3 weeks (15 sessions). The control group received no electrical stimulation. Upper limb motor function, lower limb motor function, balanceand Barthel's index were evaluated. Results In the FES group, upper limb motor function was significantly improved after treatment, and significantly better compared with the control group. Lower limb motor function showed the same significant differences. Balance was also significantly better after treatment and significantly better than in the control group. Similar significant improvements were seen in terms of Barthel's index. Conclusions FES can improve the functional capacity of elderly patients with cerebral apoplexy hemiplegia and improve their ability in daily activities.