[Novel tumor metastasis suppressorgene LASS2/TMSG1 S248A mutant promotes invasion of prostate cancer cells through increasing ATP6V0C expression].

OBJECTIVE: LASS2/TMSG1 gene is a novel tumor metastasis suppressor gene cloned from human prostate cancer cell line PC-3M in 1999 by Department of Pathology,Peking University of Basic Medical Sciences. It was found out that protein encoded by LASS2/TMSG1 could interact with the c subunit of vacuolar-ATPase (ATP6V0C). In this study, we explored the effect of LASS2/TMSG1 and its mutants on proliferation, migration and invasion of human prostate cancer cells and its molecular mechanism. METHODS: We constructed four LASS2/TMSG1 mutants and stably transfected the variants to human prostate cancer cell line PC-3M-1E8 cell with high metastatic potential. The stable transfectants were identified by qPCR and Western blot through analyzing the expression of LASS2/TMSG1 and ATP6V0C, the cell biology functions of LASS2/TMSG1 and its four mutants were studied using growth curve,MTT assay, soft agar colony formation assay, wound migration assay, Matrigel invasion study and flow cytometry. Furthermore, immunofluorescence was used to analysis the interaction of LASS2/ TMSG1 mutants and ATP6V0C. RESULTS: LASS2/TMSG1 mRNA and protein in LASS2/TMSG1 group and Mut1-Mut4 groups were higher than that in Vector group; Western blot showed that ATP6V0C protein in LASS2/TMSG1 wild group was lower than that in Vector group, but ATP6V0C protein in LASS2/TMSG1 S248A group was obviously higher than that in Vector group. MTT test and growth curve assay showed growth ability in LASS2/TMSG1 S248A group was increasing compared with other groups from day 5. Soft Agar colony formation experiment showed anchor independent growth ability in LASS2/TMSG1 S248A group was higher than those in the other groups (P<0.05), Cell migrations (from 35.3%±3.2% to 70.3%±3%) in LASS2/TMSG1 S248A group was increasing compared with LASS2/TMSG1 wild group (P<0.01), and more cells passed through Matrigel in LASS2/TMSG1 S248A group compared with LASS2/TMSG1 wild group (from 50±3.2 to 203±6.5, P<0.01), the apoptosis rate in LASS2/TMSG1 S248A group was obviously higher than that in LASS2/TMSG1 wild group (from 7% to 15.1%, P<0.05), and the G0/G1 ratio in LASS2/TMSG1 S248A group was obviously higher than that in LASS2/TMSG1 wild group (from 51.0% to 85.4%). Furthermore, double immunofluorescent staining observed the colocalization between ATP6V0C and LASS2/TMSG1 protein and its mutations, the expression of ATP6V0C in LASS2/TMSG1 S248A group increased significantly compared with the other groups. CONCLUSION: LASS2/TMSG1 S248A promotes proliferation, migration and invasion of prostate cancer cells through increasing ATP6V0C expression, suggesting that aa248-250 is an important function site for LASS2/TMSG1 in invasion suppression of prostate cancer cells.

Investigating the association between XRCC1 gene polymorphisms and susceptibility to gastric cancer.

We carried out a hospital-based case-control study to investigate the role of XRCC1 gene Arg399Gln, Arg280His, and Arg194Trp polymorphisms in susceptibility to gastric cancer. A total of 214 gastric cancer patients and 247 control subjects were recruited between March 2013 and March 2015, and polymorphism genotype frequencies were determined by polymerase chain reaction-restriction fragment length polymorphism. Using the chi-square test, we detected statistically significant differences in age (chi-square = 22.25, P < 0.001), gender (chi-square = 6.74, P = 0.01), and family history of cancer (chi-square = 4.73, P = 0.03) between the case and control groups. Logistic regression analysis revealed that the XRCC1 Arg194Trp TT genotype conferred increased susceptibility to gastric cancer compared to the CC genotype [odds ratio (OR) = 2.38, 95% confidence interval (CI) = 1.28-4.49]. Moreover, individuals carrying the T allele of this variant were found to be at moderately increased risk of this disease (OR = 1.56, 95%CI = 1.16-2.09). However, the XRCC1 Arg399Gln and Arg280His polymorphisms were shown to have no influence on the development of gastric cancer. In conclusion, we suggest that the XRCC1 gene Arg194Trp polymorphism is associated with gastric cancer susceptibility in the Chinese population.

Curative effect and safety of vascularized fibula grafting in renal transplant recipients with osteonecrosis of the femoral head: three case reports.

Osteonecrosis of the femoral head is a common and severe complication after renal transplantation. It is characterized by deterioration of hip joint function, which impairs quality of life. We present 3 renal transplant case reports of patients with osteonecrosis of the femoral head who underwent free vascularized fibular grafting at our hospital. Follow-up was from 1(1/2) to 2 years. All 3 patients exhibited good recovery with substantial improvement in joint function. Intraoperative and postoperative findings demonstrated the safety of this surgical procedure.

Dihydroetorphine-induced place preference was mediated by dopamine D1 receptors in rats.

AIM: To study the influence of dopamine (DA) receptor antagonists upon the rewarding property of dihydroetorphine (DHE). METHODS: Conditioned place preference (CPP) paradigm was used to characterize the rewarding effect of DHE. DA receptor antagonists were injected administered subcutaneously or peritoneally and microinjected into nucleus accumbens (NAcc). RESULTS: DHE (0.05, 0.5, and 5.0 micrograms.kg-1, s.c.) produced place preference (P < 0.01). Both the DA receptor antagonist haloperidol and the selective D1 receptor antagonist Sch-23390 attenuated the place preference produced by DHE (0.5 microgram.kg-1, s.c.). l-Sulpiride and spiperone, selective D2 receptor antagonists, had no such effects. CONCLUSION: The D1 (but not D2) receptors in NAcc are crucial in the mediation of the rewarding effect of DHE.

Inhibition of excessive neuronal apoptosis by the calcium antagonist amlodipine and antioxidants in cerebellar granule cells.

Neuronal cell death as a result of apoptosis is associated with cerebrovascular stroke and various neurodegenerative disorders. Pharmacological agents that maintain normal intracellular Ca2+ levels and inhibit cellular oxidative stress may be effective in blocking abnormal neuronal apoptosis. In this study, a spontaneous (also referred to as age-induced) model of apoptosis consisting of rat cerebellar granule cells was used to evaluate the antiapoptotic activities of voltage-sensitive Ca2+ channel blockers and various antioxidants. The results of these experiments demonstrated that the charged, dihydropyridine Ca2+ channel blocker amlodipine had very potent neuroprotective activity in this system, compared with antioxidants and neutral Ca2+ channel blockers (nifedipine and nimodipine). Within its effective pharmacological range (10-100 nM), amlodipine attenuated intracellular neuronal Ca2+ increases elicited by KCl depolarization but did not affect Ca2+ changes triggered by N-methyl-D-aspartate receptor activation. Amlodipine also inhibited free radical-induced damage to lipid constituents of the membrane in a dose-dependent manner, independent of Ca2+ channel modulation. In parallel experiments, spontaneous neuronal apoptosis was inhibited in dose- and time-dependent manners by antioxidants (U-78439G, alpha-tocopherol, and melatonin), nitric oxide synthase inhibitors (N-nitro-L-arginine and N-nitro-D-arginine), and a nitric oxide chelator (hemoglobin) in the micromolar range. These results suggest that spontaneous neuronal apoptosis is associated with excessive Ca2+ influx, leading to further intracellular Ca2+ increases and the generation of reactive oxygen species. Agents such as amlodipine that block voltage-sensitive Ca2+ channels and inhibit cellular oxidative stress may be effective in the treatment of cerebrovascular stroke and neurodegenerative diseases associated with excessive apoptosis.

Effect of tetrandrine on morphine dependence in isolated guinea pig ileum.

AIM: To evaluate the effects of tetrandrine (Tet) and nimodipine (Nim) on the morphine (Mor) withdrawal response in the isolated guinea pig ileum. METHODS: The withdrawal contracture was elicited by addition of naloxone (Nal) (1 mumol.L-1) to the isolated naive ileum incubated with Mor (3 mumol.L-1) at 37.5 degrees C for 4 h or to the ileum obtained from Mor-dependent guinea pig. RESULTS: When Nim (0.01, 0.05, and 0.1 mumol.L-1) or Tet (1, 10, and 50 mumol.L-1) was added 1 min before Nal in the naive ilea bathed in Krebs solution containing Mor, or when the ilea from Mor-dependent guinea pigs were incubated with Nim (0.01, 0.05, and 0.1 mumol.L-1) or Tet (1, 10, and 50 mumol.L-1) for 15 min, or when Nim (5 and 10 mg.kg-1, i.p.) or Tet (15 and 30 mg.kg-1, i.p.) was administered in vivo to Mor-dependent guinea pigs, the Nal-precipitated withdrawal contracture was significantly decreased in a dose-dependent manner. CONCLUSION: Tet and Nim, Ca2+ channel blockers, could inhibit the Nal-precipitated Mor withdrawal response in the isolated guinea pig ileum.

[Synthesis and analgesic activity of non-aryl analogues of 4-methoxycarbonyl fentanyl].

In this paper, the synthesis and analgesic activities of some derivatives of 4-methoxycarbonyl fentanyl are reported, in which the 4-N-phenyl group or both the 4-N-phenyl group and the 1-beta-phenyl group were replaced by some nonaromatic groups. In the last case, some compounds showed very strong analgesic activity. For example, the analgesic activities of compounds 4 and 6 were shown to be 695 and 818 times more potent than that of morphine respectively. The structure-analgesic activity relationships were discussed.

[Comparison of pharmacological profile of selective kappa-opioid agonist K-II and U-50488].

K-II is an analogue of U-50488, a selective kappa-opioid agonist. Comparison of pharmacological profile of K-II and U-50488 was studied using in vitro and in vivo methods. The results showed that the IC50 values of K-II and U-50488 on electrically induced contraction of the rabbit vas deferens were 0.42 nmol/L and 26.5 nmol/L, respectively. ED50 values of K-II and U-50488 in impairing motor function of mouse in the horizontal screen test were 1.7 and 15.3 mg/kg, respectively. The effect of K-II in reducing spontaneous activity of mouse was 6 times as potent as U-50488. These results suggest that K-II is a more potent kappa agonist in pharmacological effects than U-50488.

[Synthesis and analgesic activity of 1-substituted derivatives of 4-methoxycarbonyl-4-N-propionylanilinepiperidine].

In this paper, the synthesis and analgesic activities of a series of 1-substituted derivatives of N-[1-(2-phenylethyl)-4-methoxycarbonyl-4-piperidinyl]-N-propionylanil ine (4-methoxycarbonyl fentanyl) are reported. Preliminary pharmacological results showed that most compounds in this series exhibited typical morphine-like action and that replacement of beta-phenyl group in the 1-position of piperidine ring of 4-methoxycarbonyl fentanyl by some substituents e.g., some substituted vinyl groups, heterocyclic (or alcyclic) radicals, alkyl groups or N-methyl-anilino groups, can keep strong analgesic activity. Especially, some substituted vinyl groups were found to be effective groups which could replace 1-beta-phenyl group of 4-methoxycarbonyl fentanyl. Compounds N-[1-(3, 4-dimethyl-3-pentenyl)-4-methoxycarbonyl-4-piperidinyl]-N- propionylaniline(1321) and N-[1-(4-methyl-3-pentenyl)-4-methoxycarbonyl-4-piperidinyl]-N- propionylaniline(1302) exhibited higher analgesic activity than that of 4-methoxycarbonyl fentanyl.

[Effects of ohmefentanyl on rat respiration].

The respiratory depression in rats induced by sc ohmefentanyl showed a ceiling effect, but a linear relation with log dose when applied into the dorsal medulla. It elicited, however, a marked excitation on respiration when applied to dorsal pons. This excitatory effect of ohmefentanyl was antagonized by naloxone, and it antagonized to some extent the respiratory depressant effect induced by ohmefentanyl itself applied on dorsal medulla. It was postulated that the respiratory excitatory effect of sc ohmefentanyl on dorsal pons may have a bearing on the ceiling effect of its respiratory depression.